U.S. patent application number 14/202418 was filed with the patent office on 2014-10-16 for peptides for skin rejuvenation and methods of using the same.
The applicant listed for this patent is Neocutis SA. Invention is credited to Frank Dreher.
Application Number | 20140309173 14/202418 |
Document ID | / |
Family ID | 51263427 |
Filed Date | 2014-10-16 |
United States Patent
Application |
20140309173 |
Kind Code |
A1 |
Dreher; Frank |
October 16, 2014 |
Peptides For Skin Rejuvenation And Methods Of Using The Same
Abstract
The invention provides compositions for stimulating the
formation of one or more extracellular matrix components that
contain a lipoaminoacid derivative of the tripeptide carnosine such
as N-Octanoyl Carnosine. Also provided are compositions containing
N-Octanoyl Carnosine in combination with selected tripeptide and/or
tetrapeptides as well as pharmaceutical and/or cosmetic
compositions containing such compositions. The invention further
provides methods of using the compositions and compositions of the
invention to treat, alleviate, and/or ameliorate a symptom,
condition, disorder, or disease of the skin or mucosa, wherein the
symptom, condition, disorder, or disease is associated with changes
in extracellular matrix components.
Inventors: |
Dreher; Frank; (San
Francisco, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Neocutis SA |
Pully |
|
CH |
|
|
Family ID: |
51263427 |
Appl. No.: |
14/202418 |
Filed: |
March 10, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61779601 |
Mar 13, 2013 |
|
|
|
Current U.S.
Class: |
514/18.8 ;
548/339.1 |
Current CPC
Class: |
A61K 9/0019 20130101;
A61K 38/06 20130101; A61K 8/64 20130101; A61P 15/00 20180101; A61P
17/00 20180101; A61K 38/05 20130101; A61K 38/07 20130101; A61P
17/18 20180101; A61K 31/4172 20130101; A61K 8/4946 20130101; A61K
9/0014 20130101; A61K 38/07 20130101; A61K 38/05 20130101; A61Q
19/08 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61P 17/02 20180101; A61K 38/06 20130101 |
Class at
Publication: |
514/18.8 ;
548/339.1 |
International
Class: |
A61K 38/07 20060101
A61K038/07; A61K 9/00 20060101 A61K009/00; A61K 8/64 20060101
A61K008/64; A61Q 19/08 20060101 A61Q019/08; A61K 38/05 20060101
A61K038/05; A61K 38/06 20060101 A61K038/06 |
Claims
1. A composition for stimulating the formation of one or more
extracellular matrix components in skin or mucosa, comprising a
lipoaminoacid derivative of the dipeptide carnosine.
2. The composition of claim 1, wherein the lipoaminoacid derivative
of the dipeptide carnosine is N-Octanoyl Carnosine.
3. The composition of claim 2, wherein the composition further
comprises one or more carriers or excipients suitable for topical
administration or subcutaneous administration.
4. The composition of claim 3, wherein the one or more
extracellular matrix components are selected from the group
consisting of collagen I, collagen, III, collagen V, collagen VI,
collagen VII, collagen XVI, elastin, laminin, hyaluronic acid,
fibrillin, heparan sulfate proteoglycan 2, and combinations
thereof.
5. The composition of claim 4, wherein the composition stimulates
the formation of one or more extracellular matrix components
predominantly associated with skin aging selected from the group
consisting of collagen I, collagen III, collagen VII, elastin,
hyaluronic acid, and combinations thereof.
6. The composition of claim 4, wherein the composition stimulates
the formation of collagen III to a higher degree than the formation
of collagen I.
7. The composition of claim 6, wherein the composition further
comprises one or more additional active ingredients, wherein the
combination of active ingredients stimulates formation of
hyaluronic acid.
8. The composition of claim 7, wherein the one or more additional
active ingredients are selected from the group consisting of
tripeptides, tetrapeptides, and combinations thereof.
9. The composition of claim 8, wherein the composition comprises
Octanoyl Carnosine, the lipoaminoacid derivative of the tripeptide
GHK (N-Palmitoyl-GHK), and the tetra-peptide GEKG (SEQ ID
NO:1).
10. The composition of claim 3, wherein the composition is suitable
for topical administration in humans on skin, aged skin, damaged
skin, skin after cosmetic and dermatological procedures, atrophic
skin, atrophic mucosa, wounded skin, vulva, vagina, atrophic vulva,
atrophic vagina, vulvovaginal atrophy, or on skin and mucosa
associated with changes in extracellular matrix components.
11. The composition of claim 9, wherein the composition is suitable
for topical administration in humans on skin, aged skin, damaged
skin, skin after cosmetic and dermatological procedures, atrophic
skin, wounded skin, vulva, vagina, atrophic vulva, atrophic vagina,
or on skin and mucosa associated with changes in extracellular
matrix components.
12. The composition of claim 8, wherein the combination of Octanoyl
Carnosine, tripeptides, and tetrapeptides stimulates the production
of extracellular matrix components in skin or epithelial cells to a
greater degree than any one or two active ingredients alone.
13. The composition of claim 9, wherein the composition comprises
Octanoyl Carnosine, Palmitoyl-GHK, and GEKG at a weight ratio of
4:1:5.
14. The composition of claim 3, wherein the composition comprises
between about 0.0001% to 10% per weight of Octanoyl Carnosine.
15. The composition of claim 13, wherein the composition comprises
between about 0.0001% to 10% per weight of Octanoyl Carnosine.
16. The composition of 13, wherein the composition comprises about
0.004% per weight of Octanoyl Carnosine; about 0.001% per weight of
Palmitoyl-GHK, and about 0.005% per weight of GEKG, and one or more
acceptable carriers or excipients suitable for topical application
in humans on skin, aged skin, damaged skin, skin after cosmetic and
dermatological procedures, wounded skin, atrophic skin, atrophic
mucosa, atrophic vulva, atrophic vagina, or for skin and mucosa
associated with changes in extracellular matrix components.
17. A pharmaceutical composition comprising the composition of
claim 8 and one or more pharmaceutically acceptable carriers.
18. The pharmaceutical composition of claim 17, wherein the
composition comprises Octanoyl Carnosine, the lipoaminoacid
derivative of the tripeptide GHK (N-Palmitoyl-GHK), and the
tetra-peptide GEKG (SEQ ID NO:1).
19. The pharmaceutical composition of claim 18, wherein the
composition comprises Octanoyl Carnosine, Palmitoyl-GHK, and GEKG
at a weight ratio of 4:1:5.
20. A cosmetic composition comprising the composition of claim 8
and one or more cosmetically acceptable carriers.
21. The cosmetic composition of claim 20, wherein the composition
comprises Octanoyl Carnosine, the lipoaminoacid derivative of the
tripeptide GHK (N-Palmitoyl-GHK), and the tetra-peptide GEKG (SEQ
ID NO:1).
22. The cosmetic composition of claim 21, wherein the composition
comprises Octanoyl Carnosine, Palmitoyl-GHK, and GEKG at a weight
ratio of 4:1:5.
23. A kit comprising, in one or more containers, the pharmaceutical
composition of claim 18 and instructions for use.
24. A kit comprising, in one or more containers, the cosmetic
composition of claim 21 and instructions for use.
25. A method of treating, alleviating, or ameliorating a symptom,
condition, disorder, or disease of the skin or mucosa, wherein the
symptom, condition, disorder, or disease is associated with changes
in extracellular matrix components, the method comprising
administering an effective amount of the composition of claim 3 to
a patient in need thereof.
26. The method of claim 25, wherein the symptom, condition,
disorder, or disease of the skin or mucosa associated with changes
in extracellular matrix components is selected from the group
consisting of wounds, damaged skin, atrophic skin, atrophic mucosa,
aging, age-associated disorders and diseases, atrophy of any human
tissue, disorders and diseases of vulvar tissue, vulvovaginal
atrophy, and disorders and diseases of vaginal tissue.
27. The method of claim 26, wherein treating, alleviating, or
ameliorating the wound results in scarless wound healing.
28. The method of claim 26, wherein treating, alleviating, or
ameliorating atrophy of any human tissue comprises the treatment of
vulvovaginal atrophy.
29. The method of claim 10, wherein treating, alleviating, or
ameliorating skin and mucosa associated with changes in
extracellular matrix components comprises the treatment of
vulvodynia, lichen sclerosus, vulvar lichen planus, erosive lichen
planus, vulvar eczema, vulvar lichen simplex chronicus, ulcers of
the vulva, Behcet's disease, vulvar intraepithelial neoplasia, or
any combination thereof.
30. The method of claim 25, wherein the method comprises the
repeated topical administration of the composition to the
patient.
31. The method of claim 30, wherein the composition is administered
to the patient at least once or twice a day for at least 30
days.
32. The method of claim 25, wherein the method comprises the
repeated subcutaneous administration of the composition to the
patient.
33. The method of claim 32, wherein the composition is administered
to the patient at least once or twice a day for at least 30
days.
34. The method of claim 26, wherein the administration of the
composition treats, alleviates, or ameliorates one or more visible
signs of aging associated with changes in extracellular matrix
components selected from the group consisting of fine lines,
wrinkles, enlarged pores, roughness, dryness, loss of elasticity,
loss of volume, atrophic skin, atrophic vulva, atrophic vagina,
stretch marks, puffy eyes, dark (under eye) circles caused by
thinning of the skin, dark (under eye) circles caused by
insufficient blood circulation and slack tissue, and any
combinations thereof.
35. A method of improving or rejuvenating the appearance of skin
comprising administering an effective amount of the composition of
claim 3 to a patient in need thereof.
36. The method of claim 35, wherein the method results in one or
more of the following: improving the appearance of aged skin,
improving the state of aged skin, reducing the signs of aging,
preventing aging, reducing scarring of damaged skin, restoring
damaged skin, enhancing the restoration of skin after cosmetic and
dermatological procedures, restoring aging vulvar vaginal tissue,
reducing scarring of damaged skin, or restoring damaged or
pathological skin.
37. The method of claim 35, wherein the composition is administered
to the patient in the form of a cosmetic, a personal care product,
a feminine care product, a hygiene product, a dermatology product,
a pharmaceutical preparation, a medicament, or any combination
thereof.
38. The method of claim 35, wherein the composition is administered
topically to the skin or mucosa of the patient.
39. The method of claim 35, wherein the composition is administered
subcutaneously to the patient.
40. A method of maintaining healthy skin or of preventing,
ameliorating, or delaying aging of skin, the method comprising
administering an effect amount of the composition of claim 3 to a
patient in need thereof.
41. The method of claim 40, wherein the composition is administered
to the patient in the form of a cosmetic, a personal care product,
a feminine care product, a hygiene product, a dermatology product,
a pharmaceutical preparation, a medicament, or any combination
thereof.
42. The method of claim 40, wherein the composition is administered
topically to the skin or mucosa of the patient.
43. The method of claim 40, wherein the administration of the
composition prevents, ameliorates, or delays the development of one
or more signs of aging selected from the group consisting of fine
lines, wrinkles, enlarged pores, roughness, dryness, loss of
elasticity, loss of volume, atrophic skin, atrophic vulva, atrophic
vagina, stretch marks, puffy eyes, dark (under eye) circles caused
by thinning of the skin, dark (under eye) circles caused by
insufficient blood circulation and slack tissue, and any
combinations thereof.
44. A method of treating, alleviating, or ameliorating a symptom,
condition, disorder, or disease of the skin or mucosa, wherein the
symptom, condition, disorder, or disease is associated with changes
in extracellular matrix components which are not related to aging,
the method comprising administering an effective amount of the
composition of claim 3 to a patient in need thereof.
45. The method of claim 44, wherein the symptom, condition,
disorder, or disease of the skin or mucosa associated with changes
in extracellular matrix components not related to aging is selected
from the group consisting of wounded skin, damaged skin after
cosmetic and dermatological procedures, and atrophy of skin and
mucosa due to causes other than aging.
46. The method of claim 45, wherein the atrophy of skin and mucosa
is caused by emotional stress, use of oral contraceptive pills, use
of aromatase inhibitors, or any combination thereof.
47. The method of claim 44, wherein the method comprises the
repeated topical administration of the composition to the
patient.
48. The method of claim 47, wherein the composition is administered
to the patient at least once or twice a day for at least 30
days.
49. The method of claim 44, wherein the method comprises the
repeated subcutaneous administration of the composition to the
patient.
50. The method of claim 49, wherein the composition is administered
to the patient at least once or twice a day for at least 30 days.
Description
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Patent Application
No. 61/779,601, filed Mar. 13, 2013, which is herein incorporated
by reference in its entirety.
FIELD OF THE INVENTION
[0002] This invention relates generally to the fields of skin
rejuvenation of human skin, maintenance of healthy skin,
restoration of damaged skin, wound healing, treatment of atrophy of
any human tissue, and/or treatment of conditions, disorders, and
diseases of the skin and mucosa associated with changes in
extracellular matrix components. More particularly, the invention
relates to compositions and methods of using such compositions in
order to improve the appearance of aged skin by stimulating
extracellular matrix components including, for example, collagens,
elastin and hyaluronic acid in humans.
BACKGROUND OF THE INVENTION
[0003] Human skin is a complex organ which extends over the entire
body. There are different types of skin at different portions of
the body. For example, facial skin is different from that of the
scalp, and even the skin on the palm of the hand is different than
that on the back of the hand. Although the type of skin can vary
over a person's body, skin is generally composed of two main layers
of tissue. The epidermis, the outermost layer, is composed of
several layers. The dermis, also called corium or cutis vera, is
composed of a papillary layer above and a reticular layer
below.
[0004] The human epidermis is principally composed of keratinocytes
but contains also other types of cells including the melanocytes
and the Langerhans' cells. Each of these cell types contribute,
through their specific function, to the essential role played by
the skin.
[0005] The dermis provides a solid support for the epidermis. It is
also its feeder layer. The dermis consists mainly of fibroblasts
but leukocytes, mast cells or tissue macrophages are also present.
The dermis further contains blood vessels and nerve fibers. The
acellular part (i.e., the area in between the cells) of the dermis
is called extracellular matrix. The extracellular matrix of skin is
composed of various extracellular components including proteins; in
particular collagen fibers and elastin. Other extracellular matrix
components of skin include glycosaminoglycans (e.g., hyaluronic
acid, chondroitin sulfate, dermatan sulfate, keratan sulfate,
heparan sulfate, etc.), proteoglycans (e.g., fibromodulin, decorin,
biglycan, perlecan, heparan sulfate proteoglycan 2, agrin,
versican, aggrecan, lumican, collagen type IX, collagen type XII,
collagen type XIV, testican 1, testican 2, etc.) and various
glycoproteins (e.g., fibrillin 1, thrombospondin-1 and -2,
tenascin-C and -X, osteopontin, fibronectin, laminin-5 and -6,
vitronectin, etc.). These extracellular components are synthesized
by dermal fibroblasts, which make dermal fibroblasts the primary
constituent in the structural assembly of the dermis.
[0006] The extracellular matrix is a highly heterogeneous amalgam
of morphologically diverse architectural entities. It organizes and
imparts structural integrity to individual tissues, in addition to
modulating cell behavior by interacting with cell surface receptors
and soluble growth factors. Dysfunctions and changes in components
of the extracellular matrix can therefore interfere with both
tissue integrity and cell performance. Dysfunctions and changes in
components of the extracellular matrix of skin and mucosa in humans
can lead to skin aging, skin atrophy, damaged skin, wounded skin,
atrophy of vulva and vagina (vulvovaginal atrophy), or to any other
conditions, disorders and diseases of skin and mucosa associated
with changes in extracellular matrix components.
[0007] Therefore, a need exists in the art for compositions having
improved activity that maintain or even increase the level of a
rather large number of extracellular matrix components, including
those that are altered in aged, damaged, wounded, atrophic skin,
atrophy of vulva and vagina, or in any other conditions, disorders
and diseases of skin and mucosa in humans associated with changes
in extracellular matrix components.
SUMMARY OF THE INVENTION
[0008] The instant invention provides alternative peptidic
compounds (i.e., peptides such as carnosine peptidic compounds and
peptide derivatives and analogues) or appropriate peptidic
combinations thereof (i.e., combinations of peptides, peptide
derivatives and analogues), for the cosmetic and pharmaceutical
uses.
[0009] In particular, provided herein are compositions for
stimulating the formation of one or more extracellular matrix
components (e.g., collagen I, collagen III, collagen V, collagen
VI, collagen VII, collagen XVI, elastin, laminin, hyaluronic acid,
fibrillin, heparan sulfate proteoglycan 2, and/or any
combination(s) thereof).
[0010] For example, such compositions contain a lipoaminoacid
derivative of the dipeptide carnosine where the amino-terminus is
acylated. This lipoamino acid derivative may include, but is not
limited to, Octanoyl Carnosine (also referred to interchangeably
herein as N-Octanoyl Carnosine).
[0011] The chemical structure of Octanoyl Carnosine is provided
below:
##STR00001##
[0012] Octanoyl Carnosine differs in structure and biological
properties from carnosine. The chemical structure of carnosine is
provided below:
##STR00002##
[0013] Octanoyl Carnosine is a lipoaminoacid derivative of
carnosine. Octanoyl Carnosine is not naturally occurring. Rather,
Octanoyl Carnosine is a chemically modified carnosine where the
amino-terminus (--NH2) of the dipeptide carnosine has been altered
with an acylating agent to form an octanoyl group. This alteration
of the amino-terminus provides significantly different
physicochemical properties (e.g., molecular weight, diffusivity,
water solubility, lipophilicity, octanol-water distribution
coefficient, H-bond formation, etc.), chemical properties (e.g.,
chemical reactivity towards a chemical reactant, chemical
stability, spontaneous hydrolysis, transformation by carnosinase,
etc.), metabolic stability (i.e., proteolytic and/or enzymatic
degradation), and biological properties (e.g., stimulation of
extracellular matrix components including but not limited to
collagens I and III; whether or not related to different enzyme
kinetics, different affinities to cell surface or nuclear
receptors, and/or increased diffusivity through cell membranes or
other physiological barriers).
[0014] For example, being more lipophilic (i.e., higher
octanol-water distribution coefficient) than carnosine provides
Octanoyl Carnosine a higher permeability through mammal skin and
therefore a higher potency than carnosine. For example, being more
resistant to degradation (i.e., higher metabolic stability) than
carnosine provides Octanoyl Carnosine a longer and more sustained
potency than carnosine. For example, being a lipoaminoacid
derivative of carnosine provides Octanoyl Carnosine the novel and
unexpected biological properties to differently and more
effectively stimulate the formation of extracellular matrix
components; including but not limited to collagens I and III, as
compared to carnosine.
[0015] The compositions of the invention may additionally include
one or more carriers, excipients, and/or additional ingredients
suitable for topical administration and/or subcutaneous
administration.
[0016] The compositions of the invention are able to improve the
general state of skin and/or mucosa, rejuvenate skin, treat damaged
skin or mucosa, improve atrophic tissue including vulvovaginal
atrophy, and/or treat other conditions, disorders and diseases of
skin and mucosa associated with changes in extracellular matrix
components in humans.
[0017] More specifically, the invention provides peptidic
compositions, or appropriate combinations thereof, that are
sufficiently effective to be used for the stimulation of the
formation of collagen I, collagen III, collagen V, collagen VI,
collagen VII, collagen XVI, elastin, laminin, hyaluronan synthase
2, fibrillin 1, heparan sulfate proteoglycan, and/or hyaluronic
acid (or combinations thereof). Such compositions can be
particularly used for the treatment of conditions, disorders and/or
diseases of skin and mucosa associated with changes in
extracellular matrix components in humans.
[0018] Even more specifically, the compositions of the invention
stimulate the formation of one or more of the extracellular matrix
components predominantly associated with skin aging (e.g., collagen
I, collagen III, collagen V, elastin, hyaluronic acid, and/or any
combination(s) thereof). Such compositions can be particularly used
for the treatment of skin aging in humans.
[0019] Importantly, the compositions of the invention stimulate the
formation of collagen III to a higher degree than the formation of
collagen I. Determination of the degree of collagen III and/or
collagen I formation is well within the routine level of skill in
the art. Because these compositions stimulate the formation of
collagen III, they can be particularly used for the treatment of
wounded or damaged skin.
[0020] Any of the compositions of the invention can additionally
contain one or more additional active ingredients, wherein the
combination of all active ingredients stimulates the formation of
hyaluronic acid. By way of non-limiting example, the one or more
additional active ingredients may be tripeptides, tetrapeptides,
and/or any combinations thereof. Specifically, the compositions may
include Octanoyl Carnosine, in combination with the lipoaminoacid
derivative of the tripeptide GHK (N-Palmitoyl-GHK), and the
tetra-peptide GEKG (SEQ ID NO:1). Because these compositions (or
appropriate combinations thereof) stimulate the formation of
hyaluronic acid, they can be particularly used for the treatment of
wounded or damaged skin, atrophic skin and mucosa, and vulvovaginal
atrophy.
[0021] Unappreciated by the art and unexpectedly, it has been found
that compositions containing the lipoaminoacid derivative of the
dipeptide carnosine, N-Octanoyl Carnosine, stimulate the formation
of extracellular matrix components such as collagen I, collagen
III, collagen V, collagen VI, collagen VII, collagen XVI, elastin,
laminin, hyaluronic acid, fibrillin, and/or heparan sulfate
proteoglycan 2. In fact, compositions containing Octanoyl Carnosine
stimulate the formation of the extracellular matrix components
which are predominantly associated with skin aging, namely collagen
I, collagen III, collagen VII, elastin and/or hyaluronic acid.
[0022] Furthermore, unappreciated by the art and most unexpectedly,
compositions containing Octanoyl Carnosine stimulate the formation
of significantly more collagen III than collagen I. Moreover,
compositions containing Octanoyl Carnosine combined with selected
additional tri- and tetra-peptides stimulate formation of
hyaluronic acid in a synergistic manner.
[0023] Any of the compositions described herein may be suitable for
topical administration in humans on skin aged skin, damaged skin,
skin after cosmetic and/or dermatological procedures, atrophic
skin, wounded skin, vulva, vagina, atrophic vulva, atrophic vagina,
and/or on skin and mucosa associated with changes in one or more
extracellular matrix components. Moreover, in some cases, these
compositions may be suitable for subcutaneous administration in
humans.
[0024] Compositions according to the invention that contain a
combination of Octanoyl Carnosine, selected tripeptides, and
selected tetrapeptides stimulate the formation of extracellular
matrix components in skin or mucosa to a greater degree than any
one or two of the active ingredients alone. For example, the
combination of these ingredients produces synergistic results.
[0025] The invention provides compositions containing at least
Octanoyl Carnosine. Optionally, the composition may contain
Octanoyl Carnosine and at least one additionally substance (e.g.,
an acceptable carrier and/or excipient) suitable for topical
application and/or for subcutaneous application.
[0026] The invention further provides a composition containing at
least an N-acyl derivative of carnosine (other than Octanoyl
Carnosine), an ester of Octanoyl Carnosine, an ester of carnosine,
and/or any combination(s) thereof. Those skilled in the art will
recognize that any of the compositions described herein can include
Octanoyl Carnosine, any other N-acyl derivative of carnosine, an
ester of Octanoyl Carnosine, an ester of carnosine, and/or any
combination(s) thereof. These compounds are collectively referred
to herein as "derivatives of Octanoyl Carnosine" or "Octanoyl
Carnosine derivatives". The structure of suitable Octanoyl
Carnosine derivatives that can be used in the compositions of the
invention is provided below:
##STR00003##
[0027] wherein R1 is a carbon-containing side chain which can be
linear or branched, saturated or unsaturated; and can contain
heteroatoms including sulfur atoms (S), oxygen atoms (O), nitrogen
atoms (N), phosphorus atoms (P), and/or halogen atoms (F, Cl, Br,
I); and
[0028] wherein R2 is either a proton atom (H), or a
carbon-containing side chain which can be linear or branched,
saturated or unsaturated; and can contain heteroatoms including
sulfur atoms (S), oxygen atoms (O), nitrogen atoms (N), phosphorus
atoms (P), and/or halogen atoms (F, Cl, Br, I).
[0029] Preferably, the R1 group is a linear, saturated
carbon-containing side chain of --(CH2)n-CH3 whereas n is an
integer number between 0 to 18. Preferably, the R2 group is either
H, or a linear, saturated carbon-containing side chain of
--(CH2)m-CH3 whereas m is an integer number between 0 to 19.
[0030] For example for Octanoyl Carnosine, R1=--(CH2)6-CH3 and
R2=H. Octanoyl Carnosine is the most preferred derivative of
carnosine for use in the compositions and methods of the
invention.
[0031] Also provided are compositions containing at least Octanoyl
Carnosine; in combination with the lipoaminoacid derivative of the
tri-peptide GHK, N-Palmitoyl-GHK (also referred to interchangeably
herein as Palmitoyl-GHK); and the tetra-peptide GEKG (SEQ ID NO:1).
More specifically, also provided are compositions containing
Octanoyl Carnosine, in combination with Palmitoyl-GHK, GEKG (SEQ ID
NO:1), and at least one additional substance (e.g., carrier and/or
excipient) suitable for topical application.
[0032] The invention also provides compositions containing at least
Octanoyl Carnosine, an N-acyl derivative of carnosine other than
Octanoyl Carnosine, an ester of Octanoyl Carnosine, and/or an ester
of carnosine; in combination with Palmitoyl-GHK, an N-acyl
derivative of GHK other than Palmitoyl-GHK, and/or an ester
derivative of Palmitoyl-GHK; and GEKG, an N-acyl derivative of
GEKG, an ester derivative of GEKG, and/or an N-acyl GEKG ester.
[0033] Those skilled in the art will recognize that any of the
compositions described herein can include Palmitoyl-GHK, an N-acyl
derivative of GHK other than Palmitoyl-GHK, an ester derivative of
Palmitoyl-GHK, and/or any combination(s) thereof. These compounds
are collectively (and interchangeably) referred to herein as
"derivatives of Palmitoyl-GHK" or "Palmitoyl-GHK derivatives.
[0034] Likewise, those skilled in the art will also recognize that
any of the compositions described herein can include GEKG, an
N-acyl derivative of GEKG, an ester derivative of GEKG, an ester
derivative of N-acyl GEKG, and/or any combination(s) thereof. These
compounds are collectively (and interchangeably) referred to herein
as "derivatives of GEKG" or "GEKG derivatives".
[0035] In addition, those skilled in the art will also recognize
that any of the compositions described herein can also include
cyclic di-, tri, and/or tetrapeptides according to the invention.
Cyclic peptides can be obtained by either linking side chains of
the peptide or ends of the peptide sequence (head-to tail,
N-backbone to N-backbone, end to N-backbone, end to side chain,
side chain to N-backbone, side chain to side chain) through
disulfide (disulfide-bond cyclization), lanthionine, dicarba,
hydrazine or lactam bridges.
[0036] Any of the compositions described herein can also include
di-, tri, and/or tetrapeptides according to the invention where the
natural amino acid residue was substituted by unnatural amino acid
(D-configuration), an N-methyl amino-alpha-amino acid, a
non-proteogenic constrained amino acid, or a beta-amino acid.
[0037] Those skilled in the art will also recognize that any of the
compositions described herein can also include di-, tri, and/or
tetrapeptides according to the invention where the amid bond
between two amino acids is replaced by NH-amide alkylation; or as
further described in Drug Discovery Today 2010, 15, 40-56, or in
other references known in the art.
[0038] Thus, the present invention relates to compositions
containing Octanoyl Carnosine (or one or more derivatives thereof),
formulated in combination with one or more additional substances,
which can include, for example, an acceptable carrier and/or
excipient suitable for topical application and/or for subcutaneous
application. Optionally, any of the compositions described herein
may also include one or more additional substances with biological
activities.
[0039] In one aspect, the present invention provides compositions
containing Octanoyl Carnosine (or one or more derivatives thereof)
in combination with Palmitoyl-GHK (or one or more derivatives
thereof) and GEKG (or one or more derivatives thereof) at a weight
ratio of 4:1:5, formulated in combination with one or more
additional substances which can include for example, a carrier
and/or excipient suitable for topical application and/or
subcutaneous administration. Optionally, such compositions may also
include one or more additional substances with biological
activities.
[0040] Preferably, the compositions of the invention contain at
least Octanoyl Carnosine, Palmitoyl-GHK, and GEKG at a weight ratio
of 4:1:5. However, those skilled in the art will recognize that
other ratios of active ingredients can also be used and that
determination of an appropriate weight ratio can be routinely
determined.
[0041] Moreover, any of the compositions described herein can
include between about 0.0001% to 10% per weight of Octanoyl
Carnosine, depending on the solubility of Octanoyl Carnosine in the
composition. Such compositions may also contain Octanoyl Carnosine,
Palmitoyl-GHK, and GEKG (or one or more derivatives thereof) at a
weight ratio of about 4:1:5, depending on the solubilities of
Octanoyl Carnosine, Palmitoyl-GHK, and GEKG (or one or more
derivatives thereof) in the composition.
[0042] For example, some compositions according to the invention
contain about 0.004% per weight of Octanoyl Carnosine; about 0.001%
per weight of Palmitoyl-GHK, and about 0.005% per weight of GEKG
along with one or more acceptable carriers or excipients suitable
for topical administration in humans on skin, aged skin, damaged
skin, skin after cosmetic and/or dermatological procedures, wounded
skin, atrophic skin, atrophic mucosa, atrophic vulva, atrophic
vagina, and/or for skin and mucosa associated with changes in
extracellular matrix components.
[0043] The combination compositions will stimulate production of a
biomolecule whereas application of either of Octanoyl Carnosine,
the tripeptide Palmitoyl-GHK, or the tetrapeptide GEKG (or any
derivatives thereof) alone would not or would stimulate production
to a significantly lesser extent (e.g., amount) than the
combination. Alternatively (or additionally), the combination
compositions stimulate production of a greater extent of a
biomolecule than that achieved from the peptides when added or
administered individually. Indeed, in certain preferred aspects of
the present invention, the combination of Octanoyl Carnosine, the
tripeptide, and the tetrapeptide (or any derivatives thereof)
produces synergistic results.
[0044] Also provided herein are pharmaceutical and/or cosmetic
compositions and/or formulations containing any of the compositions
of the invention along with one or more pharmaceutically and/or
cosmetically acceptable carriers. In these pharmaceutical and/or
cosmetic compositions and/or formulations, the composition may
contain the combination of Octanoyl Carnosine, the lipoaminoacid
derivative of the tripeptide GHK (i.e., N-Palmitoyl-GHK) and the
tetra-peptide GEKG (SEQ ID NO:1). By way of non-limiting example,
these components of the composition and/or formulation may be
present at a weight ratio of 4:1:5.
[0045] The invention further provides kits containing, in one or
more containers, any of the pharmaceutical and/or cosmetic
compositions and/or formulations of the invention.
[0046] The compositions of the present invention are particularly
suitable for skin rejuvenation or improving the appearance of aged
skin. In addition, these compositions can also be used for
maintaining healthy skin, restoring damaged skin, enhancing the
restoration of skin after cosmetic and dermatological procedures,
wound healing including scarless wound healing, treating atrophy of
any human tissue including vulvovaginal atrophy, and for other
conditions, disorders and diseases of skin and mucosa in humans
associated with changes in extracellular matrix components.
[0047] These compositions for skin rejuvenation can be used for
improving the appearance of aged skin, maintaining healthy skin,
restoration of damaged skin, enhancing the restoration of skin
after cosmetic and dermatological procedures, wound healing
including scarless wound healing, treatment of atrophy of any human
tissue including vulvovaginal atrophy, and for other conditions,
and/or disorders and diseases of skin and mucosa in humans
associated with changes in extracellular matrix components.
[0048] Certain aspects of the present invention also relate to the
use of such compositions to make cosmetics, personal care products,
feminine care products, hygiene products, dermatology products,
pharmaceutical preparations, or medicaments for maintaining healthy
skin, skin rejuvenation, restoration of damaged skin including, but
not limited to, skin after cosmetic and dermatological procedures,
wound healing, treatment of atrophy of any human tissue including
vulvovaginal atrophy, and/or other conditions, disorders and
diseases of skin and mucosa in humans associated with changes in
extracellular matrix components.
[0049] This is accomplished by topical application of the
composition of the invention to the skin or mucosa of the human
needing such treatment. In some limited cases, this can be
accomplished by subcutaneous administration of the composition of
the invention in a human needing such treatment.
[0050] Certain aspects of the present invention also relate to
methods of using such compositions to improve the state and
appearance of human skin and to prevent and/or reduce the visible
signs of aging and/or for enhancing the restoration of skin after
cosmetic and dermatological procedures, as well as for enhancing
wound healing, reducing the atrophy of any human tissue including
vulvovaginal atrophy, and for improving other conditions, disorders
and diseases of skin and mucosa in humans associated with changes
in extracellular matrix components. These methods generally involve
topically applying the composition to the affected skin or the
affected mucosa when needed, in the amount and at the frequency
best suited for the purpose. Methods of preventing, delaying the
onset, or treating a skin or mucosal condition, disorder or disease
are also contemplated.
[0051] Furthermore the present invention relates also to the use of
Octanoyl Carnosine (or any derivatives thereof), or any peptide
combinations with Octanoyl Carnosine (or any derivatives thereof),
as active agents in medicine (i.e., active pharmaceutical
ingredients) to make a medicament.
[0052] Among the additional advantages of using peptide Octanoyl
Carnosine (or any derivatives thereof) and peptide combinations
thereof, are the fact that the peptides are less toxic in
comparison to the commonly used drugs for certain indications
mentioned herein and that the peptides have fewer side effects, can
be used for a long term treatment of conditions, disorders and
diseases of skin and mucosa in humans associated with changes in
extracellular matrix components, and can be easily administered as
composition suitable for topical application. Moreover, no toxic or
noxious degradation products of peptides are formed.
[0053] Furthermore, di-, tri-, and tetra-peptides are small (e g,
contain only two to four amino acids) and have therefore the
advantage of not being or being less immunogenic; in contrast to
larger peptides (i.e., peptides containing of five or more amino
acids) which can become immunogenic and cause allergic reactions.
This makes the di-peptide Octanoyl Carnosine, the tri-peptide
Palmitoyl-GHK and/or the tetra-peptide GEKG (or any di-, tri-, and
tetra-peptide derivatives thereof) particularly suitable for human
use.
[0054] Also, di-, tri-, and tetra-peptides are of a lower molecular
weight (grams per mol) than the peptides consisting of five or more
amino acids, which provides di-, tri-, and tetra-peptides with a
smaller molecular volume than larger peptides, which enables di-,
tri-, and tetra-peptides to better (i.e., faster, to a larger
extent) absorb and penetrate skin, mucous membranes, cell
membranes, and/or other physiological barriers. Thus, the
di-peptide Octanoyl Carnosine, the tri-peptide Palmitoyl-GHK and/or
the tetra-peptide GEKG (or any di-, tri-, and/or tetra-peptide
derivatives thereof) are particularly suitable for topical
application.
[0055] In particular, repeated topical application of Octanoyl
Carnosine (or any derivatives thereof), or preferred combinations
thereof with Palmitoyl-GHK and GEKG (or any derivatives thereof) in
accordance to the present invention can offer the advantages and
qualities described herein, as well as others which can be
routinely determined. By way of non-limiting example, these
advantages may include the ability (in some cases and with certain
preferred combinations) to improve the visible signs of aging in
human skin (including, for example, fine lines, wrinkles, skin
folds, enlarged pores, roughness, dryness, loss of elasticity, loss
of volume), improve other skin texture defects such as stretch
marks (as caused by pregnancy, trauma or other influences), reduce
bags under the eyes (also called "puffy eyes"), reduce dark (under
eye) circles (both caused by thinning of the skin, insufficient
blood circulation and slack tissue), and as well to reduce the
severity of atrophic skin and mucosa including vulvovaginal
atrophy. Moreover, some of these compositions have been discovered
to have a benefit in wound healing and tissue regeneration.
[0056] Provided herein are methods of treating, alleviating, or
ameliorating a symptom, condition, disorder, or disease of the skin
or mucosa, wherein the symptom, condition, disorder, or disease is
associated with changes in extracellular matrix components, the
method comprising administering an effective amount of any of the
compositions and/or formulations of the invention to a person
(e.g., patient) in need thereof. For example, wherein the symptom,
condition, disorder, or disease of the skin or mucosa associated
with changes in extracellular matrix components is selected from
the group consisting of wounds, aging, age-associated disorders and
diseases, atrophy of any human tissue, disorders and diseases of
vulvar tissue, disorders and diseases of vaginal tissue, and/or any
combination(s) thereof.
[0057] In some examples, treating, alleviating, or ameliorating the
wound results in scarless wound healing. In other examples,
treating, alleviating, or ameliorating atrophy of any human tissue
involves the treatment of vulvovaginal atrophy. In other examples,
treating, alleviating, or ameliorating skin and mucosa associated
with changes in extracellular matrix components comprises the
treatment of vulvodynia, lichen sclerosus, vulvar lichens planus,
erosive lichen planus, vulvar eczema, vulvar lichen simplex
chronicus, ulcers of the vulva, Behcet's disease, vulvar
intraepithelial neoplasia, and/or any combination(s) thereof.
[0058] Any of the methods described herein may involve repeated
topical administration of any of the compositions and/or
formulations described herein to the patient. By way of
non-limiting example, the compositions/and or formulations may be
administered to the patient at least once or twice a day for at
least 30 days.
[0059] Alternatively, any of the methods described herein may
involve repeated subcutaneous administration of any of the
compositions and/or formulations described herein to the patient.
By way of non-limiting example, the compositions/and or
formulations may be administered to the patient at least once or
twice a day for at least 30 days.
[0060] Those skilled in the art will recognize that the
administration of any of the compositions and/or formulations of
the invention treats, alleviates, or ameliorates one or more
visible signs of aging associated with changes in extracellular
matrix components selected from the group consisting of fine lines,
wrinkles, enlarged pores, roughness, dryness, loss of elasticity,
loss of volume, atrophic skin, atrophic vulva, atrophic vagina,
stretch marks, puffy eyes, dark (under eye) circles caused by
thinning of the skin, dark (under eye) circles caused by
insufficient blood circulation and slack tissue, and any
combinations thereof.
[0061] Those skilled in the art will further recognize that the
administration of any of the compositions and/or formulations of
the invention treats, alleviates, or ameliorates conditions,
disorders and diseases of skin and mucosa in humans associated with
changes in extracellular matrix components which are not related to
aging and which are selected from the group of wounded skin,
damaged skin after cosmetic and dermatological procedures, atrophy
of skin and mucosa due to other causes than aging (e.g., emotional
stress, use of oral contraceptive pills, use of aromatase
inhibitors, due to surgery, etc.), and for other conditions, and/or
disorders and diseases of skin and mucosa in humans associated with
changes in extracellular matrix components which are not related to
aging.
[0062] Also provided are methods of improving or rejuvenating the
appearance of skin comprising administering an effective amount of
any of the compositions and/or formulations of the invention to a
patient in need thereof. By way of non-limiting example, the
compositions and/or formulations are administered to the patient in
the form of a cosmetic, a personal care product, a feminine care
product, a hygiene product, a dermatology product, a pharmaceutical
preparation, a medicament, or any combination thereof. Such
compositions and/or formulations can be administered topically to
the skin or mucosa of the patient and/or subcutaneously to the
patient.
[0063] The invention also provides methods of maintaining healthy
skin or of preventing, ameliorating, or delaying aging of skin, the
method by administering an effect amount of any of the compositions
and/or formulations described herein to a patient in need thereof.
For example, the compositions and/or formulations may be
administered to the patient (i.e., topically to the skin or mucosa)
in the form of a cosmetic, a personal care product, a feminine care
product, a hygiene product, a dermatology product, a pharmaceutical
preparation, a medicament, or any combination thereof. In such
methods, administration of the compositions and/or formulations
prevents, ameliorates, or delays the development of one or more
signs of aging selected from the group consisting of fine lines,
wrinkles, enlarged pores, roughness, dryness, loss of elasticity,
loss of volume, atrophic skin, atrophic vulva, atrophic vagina,
stretch marks, puffy eyes, dark (under eye) circles caused by
thinning of the skin, dark (under eye) circles caused by
insufficient blood circulation and slack tissue.
[0064] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. Although
methods and materials similar or equivalent to those described
herein can be used in the practice or testing of the present
invention, suitable methods and materials are described below. All
publications, patent applications, patents, and other references
mentioned herein are incorporated by reference in their entirety.
In the case of conflict, the present specification, including
definitions, will control. In addition, the materials, methods, and
examples are illustrative only and are not intended to be
limiting.
[0065] Other features and advantages of the invention will be
apparent from the following detailed description and claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0066] FIG. 1 is a schematic showing one way of the synthesis and
purification of Octanoyl Carnosine (Octanoyl-beta-Ala-His-OH) in
its acetate salt form by solid-phase synthesis.
[0067] FIG. 2 is a schematic showing one way of the synthesis and
purification of Palmitoyl-GHK (Palmitoyl-Gly-His-Lys-OH) in its
acetate salt form by solid-phase synthesis.
DETAILED DESCRIPTION OF THE INVENTION
[0068] The present invention will be better understood from the
following description.
[0069] In this specification where reference is made to particular
features of the invention it is to be understood that the
disclosure of the invention in this specification includes all
appropriate combinations of such particular features. The
embodiments disclosed in this specification are exemplary and do
not limit the invention. As used in this specification, the
singular forms "a", "an", and "the" include plural reference unless
the context clearly dictates otherwise. The terms "comprises" and
"contains" and grammatical equivalents thereof are used in this
specification to mean that, in addition to the features
specifically identified, other features are optionally present. The
term "at least" followed by a number is used herein to denote the
start of a range beginning with that number.
[0070] This specification incorporates by reference all documents
referred to herein and all documents filed concurrently with this
specification or filed previously in connection with this
application, including but not limited to such documents which are
open to public inspection with this specification. All publications
cited herein are hereby incorporated by reference in their
entirety.
[0071] While the specification concludes with the claims
particularly pointing and distinctly claiming the invention, it is
believed that the present invention will be better understood from
the following description. The terms "having" and "including" are
to be construed as open-ended unless the context suggests
otherwise.
Extracellular Matrix Components
[0072] Collagen occurs in many places throughout the body. So far,
at least 28 types of collagen have been identified and described
which provide a variety of structural and functional properties
that collagen exhibits throughout the body. The five most common
types are collagen I, II, III, IV and V. However, over 90% of the
collagen in the body is of type I.
[0073] In human skin, collagen types I and III are the predominant
types of collagen. They are present as fibrils and are responsible
for the solidity and the strength of the dermis. Since type I
collagen is the predominant collagen in adult human skin,
comprising about 80% of the total bulk of collagen, it plays a
major role in providing tensile strength to the skin. It is clear,
however, that type III collagen, which comprises some 10% of the
total dermal collagen, also plays a critical role in providing
additional tensile properties to the skin and other tissues. (See
Journal of Dermatological Science, 24, Suppl. 1, 2000,
S60-S69).
[0074] Structurally, three collagen polypeptides wrap around each
other in a helix to form a triple helix collagen I or III molecule.
These molecules are packed in a five-stranded rope-like structure
wherein each collagen molecule is quarter-staggered with respect to
the next to form a microfibril. Microfibrils are subsequently
wrapped around other microfibrils to form fibrils, which in turn
wrap around other fibrils to produce even larger fibers.
[0075] Using histological and ultrastructural approaches in the
past, it has been well-described that chronologically aged skin
manifests in a reduced synthesis of both collagen types I and III.
With respect to photoaging, Schwarz et al. (Photochem Photobiol
1993, 58, 841-844) demonstrated that the loss in collagen in
sun-damaged human skin is due to increased degradation of both
types I and III collagens. Additionally, it was shown that
fibroblasts derived from sun-exposed skin synthesize a lower
proportion of collagen III than cells from sun-protected skin. (See
J Photochem Photobiol B. 1995, 27: 33-38).
[0076] Other collagen types are also present in skin, and a few of
them have been described to change with skin aging. For instance,
collagen VII, which is responsible for anchoring the basement
membrane onto dermal matrix, decreases with aging. (See Eur J
Dermatol 2008; 18: 297-302). One of the additional major
morphological features of aged skin is an altered dermal epidermal
junction which structurally manifests as flattening of the dermal
epidermal junction outline with the loss of rete pegs and
re-duplication of the lamina densa. Since the dermal epidermal
junction is involved in the cohesion between the dermis and
epidermis, age-related alterations in the dermal epidermal junction
as a result of collagen VII decrease entail functional changes in
skin resistance to mechanical stress and tissue homeostasis. This
may contribute to wrinkle formation.
[0077] Collagen V assembles into diverse molecular forms and
studies indicated that it is expressed in skin as different
subtypes with important but distinct roles in matrix organization
and stability. (See J Invest Dermatol 2012, 132: 1841-1849).
Whereas collagen V is the defective product in most cases of
classical Ehlers-Danlos syndrome, which is an extracellular matrix
component disorder typically characterized by skin fragility and
abnormal wound healing, it does not seem to significantly change
with skin aging.
[0078] Further collagen types such as collagens VI, XIV and XVI are
also expressed in the collagen-rich dermis. Although the structural
features of these collagens are now well-characterized, their
functions still remain mostly elusive.
[0079] The production of collagen in vivo requires activation of
the collagen biosynthesis pathway by which transcription in the
cell nucleus promotes polypeptide synthesis via translation from
mRNA, organization of the polypeptides into a pro-collagen triple
helix in the cytoplasm, secretion of pro-collagen from the cell,
and then cleavage reactions, fibril assembly, and cross-linking
extracellularly Unlike many proteins that are stored in secretory
granules and then secreted from the cell upon demand, collagen is
secreted continuously.
[0080] Alterations in content and structure of collagen and other
components of the extracellular matrix, including but not limited
to, elastin and hyaluronic acid are characteristic of aged human
skin.
[0081] Elastin is a vital component of the extracellular matrix of
vertebrates, and provides exceptional properties including
elasticity and tensile strength to many tissues and organs
including the skin. Mature elastin is an insoluble and extremely
durable protein that undergoes little turnover, but sustained
exposure to proteases may lead to irreversible and severe damage,
and thus to functional loss of the elastic fiber network. In
general, elastin content decreased with age in sun unexposed skin
(i.e., buttock) (i.e., elastin content decreased by about 44%
between 50 and 70 years of age). A similar decrease was seen in
severe sun-exposed skin (i.e., face) (i.e., elastin content
decreased by about 31% between 50 and 70 years of age).
Interestingly, the elastin content in moderately sun-exposed areas
(i.e., forearm) did not significantly change during aging. This
phenomenon might be explained by a combination of age-induced
reduction and sun-dependent increase in elastin, what appears to be
at least partially regulated by UV-induced deposition of lysozyme
in elastin fibers. (See JEADV 2006, 20, 980-987).
[0082] Fibrillins (e.g., fibrillin 1) are ubiquitous glycoproteins
of the extracellular matrix that self-polymerize into filamentous
microfibrils in which individual molecules are organized in
longitudinal head-to-tail arrays and associate laterally as well.
(See Fibrogenesis & Tissue Repair 2010, 3, 24). Fibrillin
microfibrils can additionally serve as the structural template for
tropoelastin deposition and/or crosslinking during elastic fiber
formation. Specific segments of the fibrillins interact in vitro
with numerous extracellular signaling and cell surface molecules,
including fibronectin, fibulins, syndecans and integrins. The
multiple molecular interactions of fibrillins are believed to drive
the assembly of morphologically distinct macroaggregates, which
contribute to imparting the structural integrity to individual
tissues and organs (structural role), and to target TGF-beta and
BMP complexes to the architectural matrix, which contributes to
instructing the behavior of cells (instructive role). TGF-betas and
BMPs are potent modulators of extracellular matrix metabolism that
are under the control of a complex network of relays and
servomechanisms operating within and outside the cell, and at the
cell surface. Thus, fibrillins are important components of the
extracellular matrix which are necessary for the formation of other
extracellular matrix components such as elastin and elastic fiber
formation.
[0083] Hyaluronic acid (also called hyaluronan) is an anionic,
non-sulfated glycosaminoglycan. It is unique among
glycosaminoglycans since it is non-sulfated and can be very large,
with its molecular weight (g mol-1) often reaching the millions. As
a predominant voluminous molecule, hyaluronic acid is a major
component of the extracellular matrix of the skin. It provides
structure, volume (which is associated with hyaluronic acid's
excellent water holding properties), and organization (e.g.,
facilitates the transport of ion solutes and nutrients) but also
contributes significantly to cell proliferation and migration in
the dermis. In addition, through the water-attaining properties of
hyaluronic acid, it contributes to the hydration of the skin.
[0084] Glycosaminoglycans (e.g., hyaluronic acid, chondroitin
sulfate, heparan sulfate, dermatan sulfate, keratan sulfate, etc.)
and particularly hyaluronic acid are major components of the
cutaneous extracellular matrix involved in wound healing and tissue
regeneration. Wound healing is a dynamic interactive process
involving many precisely interrelated phases, overlapping in time
and leading to the restoration of tissue integrity. The healing
process reflects the complex and coordinated body response to
tissue injury resulting from the interaction of different cell
types and extracellular matrix components. Hyaluronan plays a key
role in each phase of wound healing by stimulating cell migration,
differentiation, and proliferation as well as regulating
extracellular matrix organization and metabolism.
Glycosaminoglycans and particularly hyaluronic acid are also
involved in skin aging.
[0085] As summarized by R. Stern in 2010 (Textbook of Aging Skin,
Springer (incorporated herein by reference)), though dermal
hyaluronic acid is responsible for most hyaluronic acid in skin,
epidermal cells (e.g.; keratinocytes) are also able to synthesize
hyaluronic acid. The most dramatic histochemical change observed in
aged skin is the marked decrease in epidermal hyaluronic acid A. In
skin of an elderly person, hyaluronic acid is still present in the
dermis, while the hyaluronic acid of the epidermis has disappeared
entirely. The reasons for this precipitous fall with aging are
unknown. It has been described that the synthesis of epidermal
hyaluronic acid is influenced by the underlying dermis, thereby
indicating that epidermal hyaluronic acid is under separate
controls from dermal hyaluronic acid.
[0086] In contrast with previous in vitro and in vivo observations,
studies document that the total level of hyaluronic acid remains
about constant in human skin with aging. However, the major
age-related change is the increasing avidity of hyaluronic acid
with tissue structures with the concomitant loss of hyaluronic acid
extractability. Such intercalated hyaluronic acid may result in a
diminished ability to take on water, what results in a decreased
volume and, obviously, a loss in skin moisture. Progressive loss in
the size of the hyaluronic acid polymer in skin as a function of
age has also been reported. The increased binding of hyaluronic
acid with tissue as a function of age parallels the progressive
cross-linking of collagen and the steady loss of collagen
extractability with age. Each of these phenomena contributes to the
apparent dehydration, atrophy, and loss of volume and elasticity
that characterizes aged skin.
[0087] Other than in skin aging, collagens I and III are also major
extracellular matrix components involved in scar formation.
Scarring occurs after trauma, injury or surgery to any tissue or
organ in the body. Such scars are a consequence of a repair
mechanism that replaces the missing normal tissue with an
extracellular matrix consisting predominantly of collagen types I
and III as well as fibronectin and some other extracellular matrix
components. Scarring represents imperfect tissue regeneration.
Whereas skin wounds on early mammalian embryos (e.g., up to about
24 weeks of gestation in humans) heal perfectly with no signs of
scarring and complete restitution of the normal skin architecture,
postnatal wounds heal with scars. (See Dang C et al., Clin Plast
Surg 2003: 30, 13-23).
[0088] There are phenotypic differences between the collagen
content and cross-linking patterns in fetal and postnatal wounds
(See Clin Plast Surg 2003, 30, 13-23 and Curr Opin Pediatr 2012,
24, 371-378). In fetal wounds, type III collagen is rapidly
deposited in a fine reticular network that is indistinguishable
from uninjured skin. Post-natally, the ratio of type I to type III
collagen in wounds increases. Of the many different types of
collagen identified, fetal skin is known to contain a greater
proportion of type III collagen, whereas adult skin consists
predominantly of type I collagen. The predominance of type I
collagen in postnatal wounds provides regenerating tissue with more
strength and rigidity. Early scar formation in late gestation fetal
wounds demonstrates larger collagen fibers with greater interfiber
space.
[0089] In addition, fetal skin generally contains more hyaluronic
acid than adult skin. Further, the hyaluronic acid content of the
extracellular matrix in scarless fetal wounds is increased more
rapidly than in adult wounds. Because fetal skin contains more
hyaluronic acid than adult skin, several investigators have
therefore proposed also a role of hyaluronic acid in scarless
healing. The precise mechanisms of scarless healing remain unknown,
however, despite the great increase in knowledge gained over the
past decade.
[0090] The importance of extracellular matrix components collagen
I, collagen III, collagen V, elastin and hyaluronic acid in the
skin and the importance of maintaining, or even increasing, the
amount thereof, thus, are self-evident for skin rejuvenation and
maintaining healthy skin. In addition, collagen I, collagen III and
hyaluronic acid have been particularly recognized to be important
in wound healing and restoration of damaged skin without scar
formation.
Conditions Associated with Changes in Extracellular Matrix
Components
[0091] All terms such as "skin aging", "signs of skin aging",
"topical application", and the like are used in the sense in which
they are generally and widely used in the art of developing,
testing and marketing cosmetic and personal care products, as well
as for medicaments which are indicated for skin aging (e.g., cream
with tretinoin).
[0092] Skin aging is classified into intrinsic aging and extrinsic
aging depending on its cause. Intrinsic aging is a process by which
the skin structure and the physiological functions of the skin
deteriorate regardless of environmental changes as a human gets
older. Extrinsic aging is caused by continuous exposure to external
environment such as sunlight and air pollutants. Especially, skin
aging caused by sun light is called photoaging. Ultraviolet (UV)
light from the sun is the main cause of the physiological and
morphological changes in aged skin.
[0093] As intrinsic skin aging proceeds, the skin becomes dry, and
fine lines and wrinkles form which become more visible and deepen
with age. Further, because of structural and functional changes of
the epidermis and the dermis, the skin loses its elasticity and
looks drooping. The dermis becomes thinner and well visible skin
folds (e.g., nasolabial fold) form with age. It is estimated that
the total quantity of collagen lost each year for adults is about
1%. In addition, the remaining collagen fibers gradually become
thicker, while the cross-linking of the collagen fibers increases,
so that the solubility, elasticity and like thereof decrease.
Furthermore, elastin fibers become thicker and the cross-linking
thereof also increases. Moreover, the proliferative activity of
fibroblasts in the dermis decreases with time, and the ability of
the aging fibroblasts to form (i.e., synthesize) new collagen,
elastin, hyaluronic acid and other components of the extracellular
matrix also decreases.
[0094] Continuous exposure to the sun is the main cause of
extrinsic aging of skin. The UV component of sunlight, particularly
UVA and UVB, is generally believed to be the principal causative
agent in this process called photoaging. The extent of UV exposure
required to cause "photoaging" is not currently known, although the
amount sufficient to cause erythema (reddening, commonly described
as sunburn) in human skin is quantified as the "minimal erythema
dose" (MED) from a given UV light source. Repeated exposure to
sunlight UV at levels that cause erythema and tanning are,
nevertheless, commonly associated with photoaging.
[0095] There is a difference between the physiology of
intrinsically-aged (i.e., chronologically-aged) skin in comparison
with that of photoaged skin. Chronologically-aged skin typically
maintains a smooth and unblemished appearance, in comparison with
the leathery, blotchy, and often deep wrinkling of photoaged skin.
Photoaging is characterized clinically by coarseness, wrinkles,
mottled pigmentation, sallowness, laxity, telangiectasia,
lentigines, purpura and relative ease of bruising, atrophy,
depigmented areas, eventually premalignant, and ultimately
malignant neoplasm (i.e., abnormal mass of tissue as a result of
neoplasia, which is the abnormal proliferation of cells).
Photoaging commonly occurs in skin that is generally exposed to
sunlight such as the face, ears, bald areas of the scalp, neck,
decollete, forearms, and hands.
[0096] Although the typical appearance of photoaged and
chronologically aged human skin can be readily distinguished,
recent evidence indicates that chronologically aged and
UV-irradiated skin share important molecular features including
altered signal transduction pathways that promote
matrix-metalloproteinase expression (e.g., collagenase, gelatinase)
causing extracellular matrix degradation, decreased collagen
formation, and alteration or damage to extracellular matrix of skin
such as the accumulation of amorphous elastin-containing material
residing beneath the epidermal dermal junction. This concordance of
molecular mechanisms suggests that UV irradiation accelerates many
key aspects of the chronological aging process in human skin.
[0097] Moreover, in women after menopause, the collagen amount and
the skin thickness gradually decrease what causes a sensation of
dry or tight skin and a marked increase in skin fine lines and
wrinkles. In fact, in addition of wrinkling and loss of elasticity,
aging of the skin is also associated with skin thinning (called
atrophy) and delayed wound healing. These undesirable aging effects
are exacerbated by declining estrogen levels in postmenopausal
women. (See Am J Clin Dermatol 2011, 12, 297-311). Estrogens (e.g.,
17-beta-estradiol) stimulate keratinocyte proliferation, leading to
a thicker epidermis, and the formation of collagens and other
extracellular matrix components. Estrogen products can be therefore
used for the prevention and treatment of skin aging due to estrogen
decline with menopause. Although topical estrogen products and
systemic estrogen replacement therapy has been shown to improve
some aspects of postmenopausal skin, long-term estrogen treatment
has been associated with significant harmful systemic effects such
as increase of breast cancer rate and cardio-toxic events).
Therefore, a need for safe and effective, non-hormonal alternatives
for the treatment of post-menopausal skin exists.
[0098] Furthermore, vulvar-vaginal atrophy (also called
vulvovaginal atrophy) is another common consequence of menopause
women. (See Adv Nurse Pract 2010, 18, 31-32, 34, 55). Vulvo-vaginal
atrophy often manifests with discomfort what can be experienced as
dryness, lack of lubrication, rawness, burning, irritation,
inflammation, atrophic vaginitis, and pain. This can ultimately
lead to sexual dysfunction. Today, hormone therapy is the only
treatment approved by the US Food and Drug Administration for
vulvovaginal atrophy. Because both physicians and women are
concerned with the tolerability and safety profile of hormonal
(i.e., estrogen, estrogen plus progestin) treatments, alternative,
non-hormonal menopause therapies for the treatment of
vulvar-vaginal atrophy are needed.
[0099] Vulvovaginal atrophy occurs most often after menopause, but
it can also develop during breast-feeding, as a consequence of
breast cancer treatment, or at any other time the women's estrogen
production declines. Furthermore, recent evidence indicates that
women taking oral contraceptives (which can cause a decline in the
production of certain sex hormones such as testosterone) may also
experience vulvovaginal atrophy. (See Sex Med 2012, 9, 2213-2223;
Sex Med 2010, 7, 1585-1587). Vulvovaginal atrophy can lead to
syndromes described as vulvodynia and vestibulodynia.
[0100] Vulvodynia is a pain syndrome that affects the vulvar area
and often occurs without an identifiable cause or visible
pathology. It refers to pain of the vulva unexplained by vulvar or
vaginal infection or skin disease. Pain is the most notable symptom
of vulvodynia, and can be characterized as a burning, stinging,
irritation or sharp pain that occurs in the vulva, including the
labia and entrance to the vagina. It may be constant, intermittent
or happen only when the vulva is touched, but vulvodynia is usually
defined as lasting for months to years. Symptoms of vulvodynia may
occur in one place or the entire vulvar area. It can occur during
or after sexual activity, when tampons are inserted, or when
prolonged pressure is applied to the vulva, such as during sitting,
bike riding, or horseback riding. Some cases of vulvodynia are
idiopathic where no particular cause can be determined
Vestibulodynia, or simply vulvar vestibulitis is vulvodynia
localized to the vestibular region. It tends to be associated with
a highly localized "burning" or "cutting" type of pain. The pain of
vulvodynia may extend into the clitoris, which is referred to as
clitorodynia. Vestibulodynia is the most common subtype of
vulvodynia that affects premenopausal women--the syndrome has been
cited as affecting about 10% to 15% of women seeking gynecological
care.
[0101] Moreover, tissues from women with vulvodynia have been shown
to have a significant increase in subepithelial heparanase activity
what may lead to increased intraepithelial hyperinnervation as
compared with healthy women. (See Int J Gynecol Pathol 2008, 27,
136-141). Heparanase, which is degranulated from mast cells, is
capable of degrading the vestibular stroma and epithelial basement
membrane, thus permitting stromal proliferation and intraepithelial
extension of nerve fibers. Heparanase is an enzyme that acts within
the extracellular matrix to degrade heparan sulfate. Heparan
sulfate glycosaminoglycans (which are other examples of
extracellular matrix components of skin) are abundant components of
basement membranes and cell surfaces where they are present
associated with specific core-proteins to form proteoglycans,
mainly perlecan, glypicans and syndecans. They play many roles such
as modulation of cell proliferation and differentiation,
cell-matrix adhesion and assembly. Heparan sulfate content has been
also shown to be altered (decrease) during skin aging. This is
mainly the result of an increased formation and activity of its
degrading enzyme, heparanase (e.g., Hpse-1), due to UV-B
irradiation. (See J Photochem Photobiol B, 2012, 106. 107-112).
Heparanase activation and the consequent decrease of heparan
sulfate are associated with wrinkle formation. (See Experimental
Derm 2010, 19, 965-972).
[0102] Therefore, vulvodynia is a condition, disorder and disease
of the vulva (which is composed of both keratinized and
non-keratinized epithelial tissue) that can be also associated with
changes in extracellular matrix components.
[0103] Vulvar lichen sclerosus, a chronic inflammatory disease
which affects genital labial, perineal and perianal areas, is
another disorder associated with changes in extracellular matrix
components of the vulva. The histopathology of lichen sclerosus
suggests abnormalities in extracellular matrix composition and in
particular of proteoglycans. (See J Eur Acad Dermatol Venereol,
2012, 26, 207-212).
[0104] Vulvar lichen sclerosus produces significant discomfort and
psychological distress in peri- and post-menopausal women. Other
vulvar disorders associated with changes in extracellular matrix
components include vulvar lichen planus, erosive lichen planus,
vulvar eczema, vulvar lichen simplex chronicus, ulcers of the
vulva, Behcet's disease, and vulvar intraepithelial neoplasia.
[0105] Finally, it is also accepted that emotional stress (see
Brain Behav Immun 2009, 23, 1089-1095), tobacco smoke (see J
Investig Dermatol Symp Proc 2009, 14, 53-55), air pollution, and
certain medications (e.g., corticosteroids) (see Clin Exp Dermatol
1991, 16, 416-419) have an adverse effect on the skin and lead to
changes (e.g., decreased formation and/or increased degradation) of
collagen and other extracellular matrix components.
Compounds that Stimulate Formation of Extracellular Components
[0106] Numerous compounds including peptides that stimulate
formation of extracellular matrix components have been reported in
the scientific literature, issued patents, patent applications, or
in other communications such as technical brochures from suppliers
of those compounds. The chapter by F. Gorohhui and H. I. Maibach in
the Textbook of Aging (2010, Springer (incorporated by reference))
discloses examples of such compounds; particularly peptides.
[0107] Peptides are preferentially used for the stimulation of the
formation of extracellular matrix components for multiple reasons.
They are not toxic, or less toxic than non-peptidic compounds, and
no toxic or noxious degradation products of peptides are formed.
Thus, peptides commonly have fewer side effects, and can be used
for the long term treatment of conditions, disorders and diseases
of skin and mucosa in humans associated with changes in
extracellular matrix components. Di-, tri-, and tetra-peptides are
particularly well suited for this purpose, as they have the
advantage of not being immunogenic or of being less immunogenic, in
contrast to larger peptides which can become immunogenic and cause
allergic reactions. In addition, di-, tri-, and tetra-peptides are
of a lower molecular weight than larger peptides what enables them
to better (i.e., faster, in larger quantities) absorb and penetrate
skin, mucous membranes, cell membranes, and/or other physiological
barriers.
[0108] Furthermore, in contrast to natural extracts such as animal,
plant or vegetable extracts, peptides can be obtained of a high
purity (e.g., 95% and higher) with defined (e.g., the potential
impurities are known and their content is within the specifications
of the peptide product) and reproducible (e.g., from production
batch to production batch) characteristics. Natural extracts cannot
be obtained in a reproducible manner, frequently contain unknown
impurities which vary from batch to batch, and can contain
compounds which are not suitable for topical application since they
cause skin irritation or sensitization in susceptible individuals.
Also, natural extracts are frequently not stable and cause color
and/or odor changes of the composition containing such extracts.
This makes composition with natural extracts, which are known to
stimulate the formation of extracellular matrix components, not
useful for topical application.
[0109] The mechanisms of action of peptides are not always known.
Stimulation of the formation of extracellular matrix components may
be obtained with peptides acting, for example, on the skin's
extracellular matrix, particularly by promoting the synthesis of
molecules, by preventing their degradation, and/or by acting on
receptors.
[0110] For example, patent application WO 2007/146269 discloses
tetrapeptides that are characterized by the amino acid sequence
motif GxxG (SEQ ID NO:2) or PxxP (SEQ ID NO:3), where G (glycine)
and P (proline) residues are maintained and x is a variable amino
acid. These disclosed sequences induce production of collagen from
dermal fibroblasts as assessed by a dye-binding method designed for
the analysis of soluble collagens released into the culture medium
by mammalian cells during in vitro culture. While some of the
disclosed peptides alone or in combination induced the synthesis of
soluble collagen, none of the disclosed peptides induce the
synthesis of any specific type of collagen (i.e., such as collagen
I and/or III) or induce the synthesis of other extracellular matrix
components.
[0111] Likewise, patent application WO 2009/068351 discloses
tetrapeptides having the motifs GxxG (SEQ ID NO:2), PxxP (SEQ ID
NO:3), or PxxK (SEQ ID NO:4). Preferred peptides disclosed in this
application are tetrapeptides selected from the group of GEPG (SEQ
ID NO:5), GPPG (SEQ ID NO:6), GEKG (SEQ ID NO:1), PGPP (SEQ ID
NO:7), and/or PKEK (SEQ ID NO:8); or their N-acyl derivatives.
While some of the disclosed peptides alone or in combination induce
the synthesis of collagen I (alpha 1, or alpha 2), fibronectin 1,
or hyaluronic acid synthetase 1, none of the disclosed peptides
induce the synthesis of collagens III, hyaluronic acid, or other
extracellular matrix components.
[0112] In a subsequent clinical study (see Exp Dermatol 2011, 20,
602-604), the tetrapeptide GEKG (SEQ ID NO:1) was further shown to
stimulate hyaluronic acid formation after treatment with an
oil-in-water vehicle containing 50 ppm GEKG for sixty days.
[0113] Another example is the tripeptide Gly-His-Lys (GHK), which
is a fragment of collagen released during proteolysis of collagen,
or may be derived from the extracellular matrix binding protein
SPARC. Its copper complex (GHK-Cu) was studied for tissue
remodeling and was shown to stimulate different components of the
extracellular matrix including collagens I and III, elastin, and
some glycosaminoglycans. (See J Biomater Sci Polymer Edn 2008, 19,
969-988). An in vivo study comparing GHK with GHK-Cu in rat
experimental wounds revealed that GHK had no effect and only GHK-Cu
leads to a statistical significant increase of collagen
accumulation. (See J Clin Invest 1993, 92, 2368-2376). As shown in
this study, only low and insignificant amounts of type III collagen
were formed by GHK-Cu as compared to total collagen. Whereas higher
doses of GHK-Cu produce skin irritation (see J Biomater Sci Polymer
Edn 2008, 19, 969-988), GHK and its derivatives do not
significantly stimulate collagen III formation, or stimulate
collagen III to a significantly lesser extent than collagen I.
[0114] More recently, it was shown that the copper ion (Cu2+)
stimulates both collagen I and III, but collagen III to a lesser
extent than collagen I. (See Connective Tissue Res 2012, 53,
373-378). Therefore, the stimulation of collagen formation observed
with GHK-Cu could originate from the copper ion and not from the
GHK peptide. Copper is highly toxic and therefore not suitable for
topical or subcutaneous uses. In addition to the Cu-GHK, the
N-palmitoyl derivative of GHK (N-Palmitoyl-GHK) also stimulates
collagen synthesis. (See Int J Cosmetic Sci 2000, 22, 207-218).
[0115] Patent FR2802413 discloses that N-Palmitoyl-GHK increases
the collagen synthesis by up to 75.3% as judged from
3H-proline-incorporation experiments using human skin explants. In
these studies, the collagen increase was due to formation of
collagens I, IV and VII.
[0116] While some of these studies disclosed that GHK peptides and
derivatives thereof induce the synthesis of collagens I and III and
of a few other extracellular matrix components, none of the
disclosed peptides induce the synthesis of collagen III more than
collagen I. In fact, while GHK-Cu seems to stimulate some collagen
III synthesis, those skilled in the art will recognize that it may
be not suitable for prolonged topical application due to its
intolerability and the toxicity related to copper.
[0117] As another example, patent application WO2005/048968
discloses that selected combinations of tripeptide GHK with
tetrapeptide Gly-Gln-Pro-Arg (SEQ ID NO:9), or their analogues and
derivatives, increase the synthesis of collagens I, IV, fibronectin
and hyaluronic acid in vitro in a synergistic manner. However, none
of the disclosed peptides, analogues and derivatives, or
combinations thereof induces the synthesis of collagen III or of
collagens V and VII.
[0118] Patent application WO 2007/143006 discloses polymeric
bio-surfactants conforming to the formula Acyl-AA-Term where Acyl
is an 8- to 22-membered carbon chain and AA is a consecutive
sequence of four to nine amino acid residues, where at least one,
preferably at least two of the amino acid residues is charged, and
Term is an acid C-terminus or an amide C-terminus While some of the
disclosed polymeric bio-surfactants (alone or in combination)
induce the synthesis of collagen I, fibronectin, and elastin, none
of the disclosed compounds induces the synthesis of collagens III,
hyaluronic acid, or other extracellular matrix components.
[0119] Moreover, patent application WO2010/082175 discloses
specific peptides which significantly increase the formation of
collagen I, III, IV, fibronectin, hyaluronic acid, and laminins.
The peptide palmitoyl-KMO2K--OH increased the collagen I formation
by 111% (as determined by ELISA) and the collagen III formation by
104% (as determined by immunofluorescence) in normal human dermal
fibroblasts. Some of the disclosed peptides induce the synthesis of
both collagen I and collagen III. However, none of the disclosed
peptides induce the synthesis of collagen III more than collagen I
or induce the synthesis of collagens V and VII, and other
extracellular matrix components. In fact, also disclosed in
WO2010/082175, the addition of a specific plant extract, Portulaca
pilosa, to palmitoyl-KMO2K--OH is required to induce the synthesis
of collagen III more than collagen I by the combination in a
synergistic manner.
[0120] WO2010/136965 discloses dipeptides of the formula
R1-Tyr-Arg-R2 which significantly increase the formation of
elastin/tropelastin, fibrillin 1, and decorin in human dermal
fibroblasts. However, none of the disclosed peptides, analogues and
derivatives, or combinations thereof induces the synthesis of
collagens, or other extracellular matrix components.
[0121] Other examples known in the art are tri-peptides with the
formula K.times.K, which have been proposed as TGF-beta growth
factor activators thereby enabling the synthesis of collagen in the
skin extracellular matrix. For example, the tripeptide proposed in
FR2810323 is elaidyl-KFK. However, in patent EP1625150 the
inventors describe elaidyl-KFK as being insufficiently active to
stimulate collagen synthesis and propose to alter the lysine side
chains, for example the length of the aminated alkyl chains or to
introduce specific side chains or to use a central amino acid X
with a hydrocarbon chain possibly substituted by a hydroxyl group.
Among the examples given are palmitoyl-KVK, palmitoyl-KAK or
palmitoyl-KSK.
[0122] Also known in the art are peptides with trade names
Kollaren.RTM. (INCI name: tripeptide-1), Collaxyl.RTM. IS (INCI
name: hexapeptide-9), SYN.RTM.-TC (combination of
palmitoyl-Lys-Val-Lys-OH with palmitoyl-Lys-Val-Thr-OH and
tetradecylaminocarbonyl-Dab-Val-Dab) that significantly increase
the formation of both collagens I and III in human dermal
fibroblasts. However, none of these disclosed peptides, analogues
and derivatives, or combinations thereof induce the synthesis of
collagen III to a higher degree than the synthesis of collagen I.
In addition, those other peptides are limited to the stimulation of
only a few additional extracellular matrix components, most of
which are not associated with skin aging. Moreover, these peptides
either do not stimulate, only relatively weakly stimulate, and/or
only partially stimulate the formation of the extracellular matrix
components that have been described to be significantly altered
with skin aging, namely collagen I, collagen III, collagen V,
elastin, and hyaluronic acid. Moreover collagen III formation is
either not stimulated by the peptides of the prior art, or, it is
stimulated by the peptides to a lesser degree than collagen I
formation. Thus, in contrast to the compositions of the instant
invention, these peptides stimulate collagen I formation
significantly more than collagen III formation.
[0123] Collagen III is an important component of the extracellular
matrix. Both the formation and structure of collagen III are
changed with aging, wounding or damaging of skin, as well as many
conditions, disorders and diseases of skin and mucosa associated
with changes in extracellular matrix components. Therefore, there
is a need for compositions that effectively stimulate collagen III
formation in skin and mucosa; for compositions that effectively
stimulate collagen III without significantly stimulating collagen I
formation; and/or for compositions that effectively stimulate
collagen III significantly more than stimulating collagen I
formation. Such compositions are particularly useful for scarless
repair after wounding of skin as well as for optimal restoration of
damaged skin.
[0124] More particularly, there is a need for compositions which
stimulate collagen I, collagen III, collagen V, elastin and/or
hyaluronic acid (e.g., all of the extracellular matrix components
that are thought to be significantly altered with skin aging).
There is also a need for compositions which stimulate extracellular
matrix components such as fibrillins, which help in the formation
of elastic fibers or other extracellular matrix components.
[0125] Preferably, the compositions of the present invention
contain Octanoyl Carnosine (or one or more derivatives thereof).
The compositions of the present invention include Octanoyl
Carnosine (or one or more derivatives thereof) at concentration
sufficient for demonstrating clinical efficacy for improving skin
aging, or other skin disorders and diseases associated with skin
aging.
[0126] The compositions of the present invention incorporate
Octanoyl Carnosine (or one or more derivatives thereof) at
concentration sufficient for demonstrating clinical efficacy for
wound healing of skin, and/or for enhancing the restoration of skin
after cosmetic and dermatological procedures. The compositions of
the present invention can incorporate Octanoyl Carnosine (or one or
more derivatives thereof) at concentration sufficient for
demonstrating clinical efficacy for scarless wound healing of
wounded skin.
[0127] In another example, the compositions of the present
invention incorporate Octanoyl Carnosine (or one or more
derivatives thereof) at concentration sufficient for demonstrating
clinical efficacy for the treatment of aging vulvar and vaginal
tissue.
[0128] In another example, the compositions of the present
invention incorporate Octanoyl Carnosine (or one or more
derivatives thereof) at concentration sufficient for demonstrating
clinical efficacy for treating symptoms, disorders and diseases of
the vulvar and the vaginal tissue which can be associated with
vulvovaginal atrophy.
[0129] In yet another example, the compositions of the present
invention incorporate Octanoyl Carnosine (or one or more
derivatives thereof) at concentration sufficient for demonstrating
clinical efficacy for treating vulvodynia.
[0130] In yet another example, the compositions of the present
invention incorporate Octanoyl Carnosine (or one or more
derivatives thereof) at concentration sufficient for demonstrating
clinical efficacy for treating lichen sclerosus.
[0131] The compositions of the present invention incorporate
Octanoyl Carnosine (or one or more derivatives thereof) at
concentration sufficient for demonstrating clinical efficacy for
treating other conditions, disorders and diseases of skin and
mucosa in humans associated with changes in extracellular matrix
components, including but not limited to the treatment of atrophy
of any human tissue.
[0132] For example, the compositions of the invention contain
Octanoyl Carnosine (or one or more derivatives thereof) at a
concentration between 0.0001% to 10% Octanoyl Carnosine (or one or
more derivatives thereof) per weight; depending on the solubility
of Octanoyl Carnosine (or one or more derivatives thereof) in the
composition.
[0133] The compositions may also contain selected tri-peptides
(e.g., Palmitoyl-GHK) (or one or more derivatives thereof) and/or
selected tetra-peptides (e.g., GEKG) (or one or more derivatives
thereof). Preferably, the composition contains Octanoyl Carnosine,
Palmitoyl-GHK, and GEKG (or one or more derivatives thereof) at a
weight ratio of about 4:1:5. However, determination of other
suitable weight ratios between these components is within the
routine level of skill in the art.
[0134] When combining Octanoyl Carnosine (or one or more
derivatives thereof) with both Palmitoyl-GHK (or one or more
derivatives thereof) and GEKG (or one or more derivatives thereof),
the optimal weight ratio of Octanoyl Carnosine (or one or more
derivatives thereof) to Palmitoyl-GHK (or one or more derivatives
thereof) to GEKG (or one or more derivatives thereof) is 4 parts
Octanoyl Carnosine (or one or more derivatives thereof):1 part
Palmitoyl-GHK (or one or more derivatives thereof):5 parts GEKG (or
one or more derivatives thereof) (parts refer to parts per weight),
as discovered during in vitro studies for hyaluronic acid formation
(see Example 5, infra). Thus, one of the preferred compositions
contains Octanoyl Carnosine (or one or more derivatives thereof),
Palmitoyl-GHK (or one or more derivatives thereof) and GEKG (or one
or more derivatives thereof) at a weight ratio of 4:1:5; whereas
Octanoyl Carnosine is present in the compositions in proportions
between 0.001% and 1%, in a carrier or excipient acceptable for
topical application.
[0135] The combination of Octanoyl Carnosine, Palmitoyl-GHK and
GEKG (or one or more derivatives thereof) at the weight ratio of
4:1:5 is not known in the art. The combination of Octanoyl
Carnosine, Palmitoyl-GHK and GEKG (or one or more derivatives
thereof) at the weight ratio of 4:1:5 stimulates the synthesis of
hyaluronic acid at this specific weight ratio in synergistic and
unexpected manner.
[0136] One or more additional ingredients, including one or more
additional substances (e.g., acceptable carriers and/or excipients)
suitable for topical application can also preferably be used in
these compositions. The one or more additional ingredients may also
include additional substances with biological activities (i.e.,
biologically active agents).
[0137] Also provided are methods for improving the state and
appearance of human skin and reducing the signs of skin aging by
using a composition containing Octanoyl Carnosine (or one or more
derivatives thereof), or Octanoyl Carnosine (or one or more
derivatives thereof) in combination with one or more additional
selected tri- and/or tetra-peptides (or one or more derivatives
thereof) according to the present invention, that stimulate the
formation of extracellular matrix components.
[0138] The invention additionally provides methods of maintaining
healthy skin and/or preventing skin aging by using a composition
containing Octanoyl Carnosine (or one or more derivatives thereof),
or Octanoyl Carnosine (or one or more derivatives thereof) in
combination with selected tri- and/or tetra-peptides according to
the present invention, that stimulate the formation of
extracellular matrix components.
[0139] Additionally, the invention also provides methods of
enhancing the restoration of skin after cosmetic and dermatological
procedures, enhancing wound healing, reducing the atrophy of any
human tissue including vulvovaginal atrophy, and improving other
conditions, disorders and diseases of skin and mucosa in humans
associated with changes in extracellular matrix components by using
a composition containing Octanoyl Carnosine (or one or more
derivatives thereof), or Octanoyl Carnosine (or one or more
derivatives thereof) in combination with selected tri- and/or
tetra-peptides according to the present invention, that stimulate
the formation of extracellular matrix components.
[0140] In any of the method described herein, skin or skin cells
(e.g., epidermal keratinocytes, dermal fibroblasts) are contacted
(i.e., topically, subcutaneously, or by any other suitable method
known in the art) with a composition containing Octanoyl Carnosine
(or one or more derivatives thereof), or Octanoyl Carnosine (or one
or more derivatives thereof) in combination with selected tri-
and/or tetra-peptides. Additionally, the methods may also involve
contacting (i.e., topically, subcutaneously, or by any other
suitable method known in the art) mucosa (i.e., mucous membranes)
or mucosal cells (i.e., epithelial cells) with a composition
containing Octanoyl Carnosine (or one or more derivatives thereof),
or Octanoyl Carnosine (or one or more derivatives thereof) in
combination with selected tri- and/or tetra-peptides according to
the present invention.
[0141] The compositions can be an aerosol, emulsion, liquid,
lotion, cream, paste, ointment, foam, patch, microneedle device or
any other cosmetic, dermatological and pharmaceutically acceptable
formulation or device. Generally, an acceptable formulation for
cosmetic, dermatological, and/or pharmaceutically use would include
any acceptable carrier, excipient, and/or substance suitable for
use on human skin or mucosa. The compositions may also contain one
or more other biologically active agents including, but not limited
to, retinoids, growth factors, and/or other peptides.
[0142] Any of the compositions of the present invention may also be
used in combination with other cosmetic, skin care, feminine,
hygiene, dermatological, pharmaceutical products, and/or medical
devices.
[0143] The invention also provides methods of reducing scarring of
skin damaged by normal aging, disease, injury, trauma, or by
surgery or other medical procedures. Such methods can involve
administering to the wound of a human a composition, wherein the
composition contains any of the above-described peptides,
singularly or in combination. The compositions may also be used in
combination with other therapeutic agents, for example, such as
tissue grafts, tissue culture products, oxygen or dressings.
[0144] The compositions of the invention can be used in humans.
Alternatively, the composition may also be used in any kind of
animal, preferably in mammals, and more preferably in cows, horses,
cats, dogs, pigs, goats, or sheep.
Carnosine
[0145] As used herein, the term "carnosine" includes and
encompasses the di-peptides beta-alanyl-histidine (see Formula II,
supra) and all related compounds such as anserine
(beta-alanyl-1-methyl-histidine) and homocarnosine
(gamma-amino-butyryl-histidine). As used herein the term
"carnosine" also includes D, L-carnosine, D-carnosine, L-carnosine,
as well as salts thereof.
[0146] Carnosine's properties, functions and potential therapeutic
applications have been extensively reviewed. (See P. J. Quinn et
al. (Molec Aspects Med 1992, 13, 379-444) (incorporated herein by
reference)). Although carnosine's anti-aging properties have
previously been described, the properties of Octanoyl Carnosine
have not previously been described and are surprising and
unexpected. As reviewed by A. R. Hipkiss (Int J Biochem Cell Biol
1998, 30, 863-868; Cell Mol Life Sci 2000, 57, 747-753;
Experimental Gerontology 2009, 44, 237-242 (incorporated herein by
reference)), the naturally occurring dipeptide
beta-alanyl-L-histidine (L-carnosine) is found in large amounts in
long-lived tissues. It has protective functions in addition to its
anti-oxidant and free-radical scavenging roles, and it extends
cultured human fibroblast life-span, kills transformed cells,
protects cells against aldehydes and an amyloid peptide fragment,
and acts as an anti-glycating agent. More recently, carnosine was
shown to protect against telomere shortening in cultured human
fibroblasts and to extend the life-span of senescence-accelerated
mice and Drosophila flies. Studies have revealed carnosine-induced
upregulation of stress protein expression and nitric oxide
synthesis, both of which may stimulate proteasomal elimination of
altered proteins. As carnosine exerts anti-convulsant effects in
rodents, the dipeptide may participate in the repair of protein
isoaspartyl groups.
[0147] Carnosine has also been shown to promote healing of skin
wounds, gastric and duodenal ulcers, as well as corneal and
pulmonary wounds. (See Nutrition 1998, 14, 266-269; Molec Aspects
Med 1992, 13, 379-444). For instance, it has been shown to
accelerate healing of bleomycin-induced and irradiation-induced
pulmonary wounds. (See Am J Physiol Lung Cell Mol Physiol 2007,
292, L1095-L1104; Med Hypotheses 2006, 66, 957-959). These effects
of carnosine have been attributed to its affinity to quench free
radicals. In dermal wounds, carnosine promotes granulation,
increases the tensile strength and hydroxyproline content in the
wound area. (See Surgery 91:56-60 (1982); Surgery 100:815-21
(1986)). The observed effect has been ascribed to the histamine
synthesis from histidine, one of the components of carnosine and
the stimulation of collagen synthesis by beta-alanine, another
component of carnosine.
[0148] Other investigations on the role of carnosine in the wound
healing of surgical wounds have shown that carnosine is enhancing
biosynthesis of glycosaminoglycans. (See Surgery 91:56-60 (1982);
Cell Mol Biol Inc Cyto Enzymol 23:267-73 (1978); Cell Mol Biol
29:1-9 (1983)). Nagai et al. (Surgery 100: 815-821 (1986)) explain
the mechanisms of action of carnosine in wound healing as
beta-alanine induced collagen synthesis combined with histamine
formation, which promotes granulation and results in a faster
healing of the wound tissue.
[0149] In a more recent study of diabetic wound healing in a
diabetic animal model (see Amino Acids 2012, 43:127-134), it was
observed that daily injections and local application of carnosine
significantly enhanced the expression of extracellular matrix
components collagen I and smooth muscle actin, as well as certain
growth factors. Carnosine treatment caused a significant increase
in IGF1 expression in the wound area suggesting a direct or
indirect modulation of IGF1 expression by carnosine enabling
improved wound healing. Similarly, TGF-beta, which is well studied
for its role in cutaneous wounds in various phases of wound
healing, is significantly over-expressed in the wound tissue from
carnosine-treated.
[0150] Apart from these effects, the wound healing potential of
carnosine may also be due to its ability to alter the generation of
free radicals and oxidative stress-induced prolonging of the
inflammatory phase of the wound healing process in a diabetic
wound. Since carnosine also shifts the acid-base balance to higher
pH values, it can modify the wound micro-environment and block the
activity of acid proteases and thereby enhance wound healing under
hyperglycemic conditions.
[0151] In another recent study (see Neuro Endocrinol Lett 2010, 31
Suppl 2: 96-100), carnosine was shown to inhibit the degradation of
hyaluronan induced by free radicals (hydroxy and peroxy-type
radicals) in vitro. Importantly, as demonstrated in this study, and
in contrast to the findings of the present invention using Octanoyl
Carnosine (or one or more derivatives thereof) and combinations of
Octanoyl Carnosine (or one or more derivatives thereof) with
selected tri- and/or tetra-peptides, carnosine does not induce the
formation of hyaluronan but, rather, limits its degradation through
carnosine's antioxidant properties. Thus, the increase in
glycosaminoglycans observed in earlier studies (see, e.g., Surgery
91:56-60 (1982); Cell Mol Biol Incl Cyto Enzymol 23:267-73 (1978);
Cell Mol Biol 29:1-9 (1983)) is the result of a decreased
degradation and not of an enhanced formation of
glycosaminoglycans.
[0152] Carnosine was also shown to inhibit high-glucose-mediated
matrix accumulation in human mesangial cells by interfering with
TGF-.beta. production and signaling. (See Nephrol Dial Transplant
2011, 26, 3852-3858). Under high-glucose conditions, deposition of
collagen VI and fibronectin were increased, what was significantly
inhibited on the protein and messenger RNA level by carnosine.
TGF-.beta. production increased under high-glucose conditions but
was completely normalized by addition of L-carnosine.
[0153] Moreover, in a study investigating liver disorders induced
by Schistosoma mansoni parasite in hamsters, subcutaneous
administration of carnosine lowered the serum pro-collagen III
peptide level in infested hamsters. (See Comp Biochem Physiol B
Biochem Mol Biol 2002, 131, 535-542).
[0154] In diabetic nephropathy, carnosine inhibited the increased
formation of fibronectin and collagen type VI in podocytes and the
increased production of TGF-beta in mesangial cells induced by
glucose. (See Diabetes 2005, 54, 2320-2327). Thus, carnosine
protects against the adverse effects of high glucose levels on
renal cells.
[0155] Therefore, all these effects (including carnosine's wound
healing properties) can be attributed to carnosine's anti-oxidant
properties, anti-glycation effects (in particular under
hyperglycemic conditions), as well as the histamine synthesis from
histidine as a component of carnosine, and/or stimulation of
collagen I synthesis by beta-alanine, which is another component of
carnosine.
[0156] In summary, carnosine has not been described to induce the
formation of collagen III to a higher degree than collagen I, or to
stimulate the formation of all extracellular matrix components that
have been described to be significantly altered with skin aging,
namely collagen I, collagen III, collagen V, elastin, and
hyaluronic acid.
Octanoyl Carnosine and Other N-Acylated Derivatives of
Carnosine
[0157] The term "N-Octanoyl Carnosine" (also referred to
interchangeably herein as "Octanoyl Carnosine") in accordance with
the present invention includes and encompasses all derivatives of
carnosine where the amino-terminus of the di-peptide is acylated to
form an octanoyl group. As used herein the term "Octanoyl
Carnosine" includes octanoyl D,L-carnosine, octanoyl D-carnosine,
octanoyl L-carnosine; as well as their salts. The terms Octanoyl
Carnosine and N-Octanoyl Carnosine are synonyms that are used
interchangeably herein. Octanoyl Carnosine can be produced by
reaction of carnosine with octanoic acid (caprylic acid) under
appropriate conditions. (See Example 1, infra).
[0158] Octanoyl Carnosine is an example of an N-acylated derivative
of carnosine. N-Acylated derivatives of carnosine can be obtained
after reacting the free amino-group (N-terminal amino group) of
carnosine with an acylating agent under appropriate conditions
known in the art of peptide synthesis. The acylating agent can be
activated, using general techniques known in the art such as those
described for example in "Amide bond formation and peptide
coupling" (Tetrahedron 61(46), 10827-10852, 2005). Examples of
activated acylating agents include, but are not limited to, acid
chlorides, acid bromides, acid fluorides, symmetrical anhydrides,
mixed anhydrides, carboxylic acids activated using common
carbodiimides such as, but not limited to, diisopropylcarbodiimide
(DIPCDI), N,N'-dicyclohexylcarbodiimide (DCC),
1-ethyl-3-(3'-dimethylamino-propyl)carbodiimide hydrochloride
(EDC). Other non-limiting examples include carboxylic acids using
the aforementioned carbodiimides and an additive, including, but
not limited to, N-hydroxysuccinimide (HOSu), N-hydroxybenzotriazol
(HOBt), 1-Hydroxy-7-azabenzotriazol, 6-chloro-N-hydroxybenzotriazol
(HOAt), 3-Hydroxy-3,4-dihydro-4-oxo-1,2,3-benzotriazine (DhbtOH) or
p-nitrophenol (PNP). Other examples include, but are not limited
to, carboxylic acids activated with an uronium salt or a
phosphonium salt, such as but not limited to,
O-Benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate
(HBTU), O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
tetrafluoroborate (TBTU),
2-(6-Chloro-1H-benzotriazole-1,1,3,3-tetramethylaminium
hexafluorophosphate (HCTU),
2-(6-Chloro-1H-benzotriazole-1-yl)-1,1,3,3-tetramethylaminium
tetrafluoroborate (TCTU),
2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate (HATU),
2-(3,4-Dihydro-4-oxo-1,2,3-benzotriazin-3-yl)-N,N,N',N'-tetramethyluroniu-
-m hexafluorophosphate (HDBTU),
2-Succinimido-1,1,3,3-tetramethyluronium hexafluorophosphate
(HSTU), N,N,N',N'-Tetramethyl-O-(succinimidyl)uronium
tetrafluoroborate (TSTU),
2-(endo-5-norbornene-2,3-dicarboxymido)-1,1,3,3-tetramethyluronium
hexafluorophosphate (HNTU),
1-benzotriazolyoxytris-(dimethylamino)phosphonium
hexafluorophosphate (BOP) or
benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate
(PYBOP).
[0159] Other activated acylating agents include, but are not
limited to, esters of N-hydroxysuccinimide (NHS ester),
p-nitrophenol (PNP ester),
N-Hydroxy-5-norbornene-2,3-dicarboxylimide (HONB-ester),
N-pentafluorophenol ester (PfP-ester), 2,4-dinitrophenyl ester,
4-nitrophenyl ester,
3-hydroxy-3,4-dihydro-4-oxo-1,2,3-benzotriazine (HODhbt),
carbonyldiimidazole (CDl) or
N-ethyl-5-phenylisoxazolium-3'-sulfonate (NEPIS), preferably a
N-hydroxysuccinimide ester, p-nitrophenol or a HOBt ester or a
derivative thereof using reaction conditions as, e.g., described in
Organic Synthesis on solid Phase (Florencio Zaragoza Dorwald,
Wiley-VCH Verlag GmbH, D-69469 Weinheim, 2000), Novabiochem Catalog
(Merck Biosciences 2006/2007) and Fmoc Solid Phase Peptide
Synthesis (Edited by W. C. Chan and P. D. White, Oxford University
Press, 2000, ISBN 0-19-963724-5).
[0160] As known in the art, the same (or similar) methods of
obtaining or producing N-acylated derivatives of carnosine can also
be used to obtain N-acylated derivatives of other peptides,
including but not limited to, the tri-peptide GHK, GHK esters, the
tetra-peptide GEKG, and GEKG esters.
N-Acetyl-Carnosine
[0161] The N-acetylated derivative of carnosine, N-acetylcarnosine,
has previously been described for uses in ophthalmology. (See
Recent Pat Drug Deliv Formul 2009, 3, 229-65). For instance, patent
applications WO 2004/028536 A1; WO 94/19325; WO 95/12581; WO
2004/064866 A1 describe N-acetylcarnosine lubricant eye drops
designed as 1% N-acetylcarnosine prodrug of L-carnosine containing
a mucoadhesive cellulose-based compound combined with corneal
absorption promoters in a drug delivery system for the management
of age-related serious or disabling eye diseases in humans
(age-related cataracts, ocular inflammation, age-related macular
degeneration, macular dystrophies, ocular manifestations of
diabetes, hypertonic retinopathy, primary open angle glaucoma,
vitreous lesions).
[0162] Acetylation makes N-acetylcarnosine more resistant to
degradation by carnosinase, an enzyme that breaks down carnosine to
its constituent amino acids, beta-alanine and histidine. This
results in the prolongation and potentiation of physiological
responses to treatments with carnosine as antioxidant and
anti-glycation agent. (See Life Sci 2006, 11, 78, 2343-57). For the
prolongation and potentiation of the antioxidant protective effects
to the cosmetic, therapeutic and medical treatments with carnosine,
N-acetylcarnosine has also been proposed for the use in skin care
products. (See Life Sci 2006, 78, 2343-2357).
[0163] As is true for N-acetylcarnosine, Octanoyl Carnosine is also
highly resistant to hydrolysis by carnosinases and provides the
antioxidant protective and anti-glycation effects characteristic
for carnosine for a longer period of time as compared to unmodified
carnosine.
[0164] N-acetylcarnosine has not been described to induce the
formation of collagen III to a higher degree than collagen I or to
stimulate the formation of all extracellular matrix components that
have been described to be significantly altered with skin aging,
namely collagen I, collagen III, collagen V, elastin, and
hyaluronic acid.
Esters of Carnosine
[0165] Esters of carnosine include ester derivatives of carnosine
which can be obtained after reacting the free carboxyl-group (free
acid C-terminus) of carnosine with an esterifying agent under
appropriate conditions known in the art of peptide synthesis. In
one non-limiting method, an acidified alcohol solution is used for
the esterification of carnosine. The method includes, for example,
the use of an acidified alcohol such as methanolic HCl to generate
a carnosine ester, e.g., carnosine methyl ester. Additionally,
other alcohols (such as, for example, ethanol, propanol, or
isopropanol) or substituted alcohols (such as, for example,
aminoethanol) can be used to generate a reagent for esterification
of the free acid C-terminus of carnosine.
[0166] Suitable alcohols for esterification of carnosine include
but are not limited to organic chemicals with at least one hydroxyl
group such as alkyl alcohols (e.g., C1 to C20 primary alcohols, C3
to C20 secondary alcohols), aryl alcohols (e.g.: benzyl alcohol),
or poly-alcohols such as sugar alcohols (e.g., mono-, di-,
poly-saccharides) or glycols and poly-glycols (e.g., glycerin,
ethylene glycol, propylene glycol, PEGs, PPGs). Ascorbic acid is
also a suitable agent (since it has at least one free hydroxyl
group) useful for esterification with carnosine.
[0167] Any methods to obtain esters of carnosine known in the art
can also be used to obtain esters of other peptides (including the
esterification of free carboxyl groups of the peptides other than
the free acid C-terminus) including, but not limited to, peptides
GEKG, N-Acyl-GEKG, and/or N-Palmitoyl-GHK.
[0168] For example, PEGylation of carnosine, Octanoyl Carnosine,
N-Palmitoyl-GHK,
[0169] GEKG, and/or N-Acyl-GEKG is one preferred esterification
method to prolong half-life, increase stability and provide higher
water solubility of those peptides.
Peptides
[0170] The term "peptide" in accordance with the present invention
is a compound that includes an uninterrupted sequence of at least
two amino acids within its structure. The terms "di-peptide" or
"dipeptide" as used herein refer to a compound that includes an
uninterrupted sequence of two amino acids within its structure. The
terms "tri-peptide" or "tripeptide" as used herein refer to a
compound that includes an uninterrupted sequence of three amino
acids within its structure. As used herein, a "tetra-peptide" or
"tetrapeptide" is a compound that includes an uninterrupted
sequence of four amino acids within its structure. These amino
acids are indicated herein using a traditional one letter
convention from left (N-terminal end) to right (C-terminal end). In
this nomenclature, G is glycine, H is histidine, K is lysine, and E
is glutamic acid.
[0171] The term "amino acid" as used herein includes and
encompasses all of the naturally occurring amino acids, either in
the D- or L-configuration if optically active, and the known
non-native, synthetic, and modified amino acids, such as
homocysteine, ornithine, norleucine and p-valine. A list of
non-natural amino acids may be found in The Peptides, Vol. 5
(1983), Academic Press, Chapter VI, by D. C. Roberts and F.
Vellaccio (incorporated herein by reference). The amino acids in
the peptides of the present invention may be present in their
natural L-configuration, unnatural D-configuration, or as a racemic
mixture.
[0172] As used herein, the term "peptide" shall also refer to
salts, deprotected forms, acylated forms of the peptide, deacylated
forms of the peptide, enantiomers, diastereomers, racemates,
prodrugs and hydrates of the above-mentioned peptide. Diastereomers
of the peptide are obtained when the stereochemical or chiral
center of one or more amino acids is changed. The enantiomer has
the opposite stereochemistry at all chiral centers.
[0173] The term "prodrug" refers to any precursor compound which is
able to generate or to release the above-mentioned peptide under
physiological conditions. Such prodrugs are for instance larger
peptides which are selectively cleaved in order to form the peptide
of the invention. Further prodrugs are protected amino acids having
especially protecting groups at the carboxylic acid and/or amino
group. Suitable protecting groups for amino groups are the
benzyloxycarbonyl, t-butyloxycarbonyl (BOC), formyl, and acetyl or
acyl group. Suitable protecting groups for the carboxylic acid
group are esters such as benzyl esters or t-butyl esters.
[0174] Peptides are synthesized by coupling the carboxyl group or
C-terminus of one amino acid to the amino group or N-terminus of
another. Due to the possibility of unintended reactions, protecting
groups are usually necessary. Chemical peptide synthesis starts at
the C-terminal end of the peptide and ends at the N-terminus.
Peptides can be synthesized either by solid-phase peptide
synthesis, by liquid-phase peptide synthesis, or by fragment
condensation. In principle, the seemingly simple formation of a
peptide bond can be accomplished using all the procedures available
in organic chemistry for the synthesis of carboxylic acid amides.
However, due to the presence of various functional groups in
natural and unnatural amino acids and particularly the requirement
for full retention of chiral integrity, the coupling of amino acids
and peptides under mild conditions can be challenging. A plethora
of coupling reagents has been developed suitability for specific
applications (e.g., solid-phase peptide synthesis or fragment
condensation). All coupling methods have the same reaction
principle in common--after activation of the carboxyl group of the
first amino acid, the second amino acid can form the peptide bond
by a nucleophilic attack of its amino group. In order to prevent
uncontrolled peptide bond formation the amino group of the first
amino acid and all functional side chain groups need to be
reversibly blocked. Repeated de-blocking, activation, and coupling
build the peptide to its desired final sequence.
[0175] The general process for synthesizing peptides on solid-phase
(e.g., resin) starts by attaching the first amino acid, the
C-terminal residue, to the resin. To prevent the polymerization of
the amino acid, the alpha amino group and the reactive side chains
are protected with a temporary protecting group. Once the amino
acid is attached to the resin, the resin is filtered and washed to
remove byproducts and excess reagents. Next, the N-alpha protecting
group is removed in a deprotection process and the resin is again
washed to remove byproducts and excess reagents. Then the next
amino acid is coupled to the attached amino acid. This is followed
by another washing procedure, which leaves the resin-peptide ready
for the next coupling cycle. The cycle is repeated until the
peptide sequence is complete. Then typically, all the protecting
groups are removed and the peptide resin is washed, and the peptide
is cleaved from the resin.
[0176] The invention is further directed towards a method for
producing the disclosed peptides. The peptides may be produced
using any method known to those skilled in the art such as those
disclosed in Merrifield, R. B., Solid Phase Peptide Synthesis I.,
J. AM. CHEM. SOC. 85:2149-2154 (1963); Carpino, L. A. et al.,
[(9-Fluorenylmethyl)Oxy]Carbonyl (Fmoc) Amino Acid Chlorides:
Synthesis, Characterization, And Application To The Rapid Synthesis
Of Short Peptides, J. ORG. CHEM. 37:51:3732-3734; Merrifield, R. B.
et al., Instrument For Automated Synthesis Of Peptides, ANAL. CHEM.
38:1905-1914 (1966); or Kent, S. B. H. et al., High Yield Chemical
Synthesis Of Biologically Active Peptides On An Automated Peptide
Synthesizer Of Novel Design, IN: PEPTIDES 1984 (Ragnarsson U., ed.)
Almqvist and Wiksell Int., Stockholm (Sweden), pp. 185-188, all of
which are incorporated by reference herein in their entirety.
Preferably, the peptides are manufactured by solid-phase synthesis.
However, the peptides may also be manufactured using standard
solution phase methodology.
[0177] Preferably, the peptides are in their trifluoroacetate (TCA)
and/or acetate salt forms. However, the peptides can be in any
other salt form including, but not limited to, adipate, ascorbate,
alginate, benzoate, benzenesulfonate, bromide, carbonate, citrate,
chloride, dibutyl phosphate, dihydrogen citrate, dioctyl phosphate,
dihexadecyl phosphate, fumarate, gluconate, glucuronate, glutamate,
hydrogen carbonate, hydrogen tartrate, hydro-chloride, hydrogen
citrate, iodide, lactate, liponic acid, malate maleate, malonate,
palmoate (embonate), palmitate, phosphate, salicylate, stearate,
succinate, sulfate, tartrate, tannate, oleate, octyl phosphate; any
other salts of the phosphate or carboxylate family; and/or any
combination thereof. Under certain conditions, other salts can be
derived from inorganic bases, include by way of example only,
sodium, potassium, lithium, ammonium, calcium and magnesium salts.
Salts derived from organic bases include, but are not limited to,
salts of primary, secondary and tertiary amines.
[0178] The term "peptide", as used herein, includes proteins.
Skin Aging
[0179] "Signs of skin aging" include, but are not limited to, all
outward visibly and tactilely perceptible manifestations as well as
any other macro or micro effects due to skin aging. Such signs may
be induced or caused by intrinsic factors (showing as chronological
aged skin) and extrinsic factors (showing as environmental skin
damage including but not limited photo-aged skin). These signs may
result from processes which include, but are not limited to, the
development of textural discontinuities such as wrinkles and coarse
deep wrinkles, fine or skin lines, crevices, bumps, large pores
(e.g., associated with adnexal structures such as sweat gland
ducts, sebaceous glands, or hair follicles), or unevenness or
roughness, loss of skin elasticity (loss and/or inactivation of
functional skin elastin), sagging (including puffiness in the eye
area and jowls), loss of skin firmness, loss of skin tightness,
loss of skin recoil from deformation, discoloration (including
under eye circles), blotching, sallowness, hyperpigmented skin
regions such as age spots and freckles, keratoses, abnormal
differentiation, hyperkeratinization, elastosis, collagen
breakdown, and other histological changes in the stratum corneum,
dermis, epidermis, the skin vascular system (e.g., telangiectasia
or spider vessels), and underlying tissues, especially those
proximate to the skin.
[0180] As used herein, prophylactically regulating a skin condition
includes delaying, minimizing and/or preventing visible and/or
tactile discontinuities in skin (e.g., texture irregularities in
the skin which may be detected visually or by feel), including
signs of skin aging. As used herein, therapeutically regulating
skin condition includes ameliorating, e.g., diminishing, minimizing
and/or effacing, discontinuities in skin, including signs of skin
aging. Some of the compositions of the present invention may be
used for prophylactically and/or therapeutically regulating a skin
condition.
[0181] Some of the compositions of the present invention are useful
for improving skin appearance and/or feel. For example, some
preferred compositions of the present invention are useful for
regulating the appearance of skin condition by providing an
immediate visual improvement in skin appearance following
application of the composition to the skin. Generally speaking,
compositions of the present invention which further contain
particulate materials will be most useful for providing the
immediate visual improvement.
Demonstrating Clinical Efficacy
[0182] Prevention, amelioration, and/or treating of the signs of
skin aging, protection and/or improving skin condition, and the
prevention and/or treatment of skin imperfections are functional
features which can be visualized, analyzed, measured and quantified
using many techniques known by the specialist in cosmetic or skin
rejuvenation treatments. Decrease of fine lines, wrinkles, skin
folds, and of skin roughness can be quantified either directly on
the person contact-free using fringe projection (FOITS=Fast Optical
In vivo Topometry System; Dermatop.TM. or Primos.TM. system), or by
silicon replicas of the skin area which are then analyzed by the
technique called "drop shadows" or by a FOITS system, or by a
Canfield VISIA.TM. device. Changes in volume and shape of the face
can be quantified using a relief obtaining system without contact
using a fringe projection FOITS system. Alteration of the skin
barrier can be quantified by measuring transepidermal water loss
(TEWL) using a Tewameter.TM., a Vapometer.TM., a Dermalab.TM.,
and/or an Aquaflux.TM. device. Loss of firmness and/or elasticity
and/or tone and fatigue of the skin can be quantified using a
Cutometer.TM., a Reviscometer.TM., an Aeroflexmeter.TM., a
Dynaskin.TM., a Ballistometer.TM., a Twistometer.TM. and/or a
Dermalab.TM. device. Dull complexion, loss of uniformity of skin
tone, pigmentation changes (hypo and hyper pigmentation), local
reddening, loss of clarity and brightness of the complexion,
pigmentation spots, rosacea, dark circles are directly measurable
using a Mexameter.TM., a Chromameter.TM., a Colormeter.TM., a
Canfield VISIA.TM., a Canfield VISIA-CR.TM., a SIAscope.TM., a
Goniolux.TM. or a confocal laser microscope device, and/or by
specific color analysis on photo (enabled by the technique of
photographing in polarized crossed and parallel light). The number
and size of facial pores can be quantified by the silicon replica
technology described above, or by specific analysis on photo
(enabled by using a video microscope or a macroscopic photographing
system). Atrophy and thinning of the skin, epidermis, dermis, or
hypodermis (e.g., in case of studying slimming agents) is
measurable by measuring TEWL (e.g., in case of studying the
epidermis), or by an ultrasound echographic device, and/or a
confocal laser microscope device. Density of skin fibers can be
quantified by ultrasound and then by image analysis. Cellulite is
quantified either directly by a relief obtaining system without
contact using fringe projection (FOITS) or indirectly by measuring
the length of the dermo-hypodermal junction by an ultrasound
echographic device. Stretch marks are either directly quantified
using a relief obtaining system without contact using fringe
projection (FOITS) or by the silicon replica technology. Skin
softness is directly measurable by techniques of friction study as
with a frictiometer device or indirectly by the silicon replica
technology. Changes in collagen, extracellular matrix components,
and/or in connective tissue fibers may be quantified by histology,
confocal laser microscopy, UV spectroscopy, SIAscopie, and/or by
multiphoton spectroscopy. All changes visible to the eye (including
but not limited to fine lines, wrinkles, folds, texture, sagging,
loss of elasticity color, tone, pigmentation, redness) can be
quantified in direct or on photography, by a trained judge person
or not, with or without visual scoring system (e.g., using a
4-point severity scale).
Cosmetic Composition and Medicament
[0183] The terms "cosmetic composition" and "cosmetic product" are
used interchangeably herein relate formulations that can be used
for cosmetic purposes or purposes of hygiene or as a basis for
delivery of one or more cosmetic and/or pharmaceutical substances,
products, and/or ingredients.
[0184] The terms "pharmaceutical composition" and "medicament" is
used herein to refer to a formulation that can be used for medical
purposes or as a basis for delivery of one or more cosmetic and/or
pharmaceutical substances, products, and/or ingredients.
[0185] It is possible that any of the formulations, compositions,
medicaments, and/or products described herein can be used for two
or more of these same purposes at one time.
[0186] Preferably, the compositions described herein are suitable
for "topical application" (i.e., on top of skin surface, on top of
mucosal surface). As used herein, topical application includes, but
is not limited to, cutaneous; ocular; mucosal; buccal; vaginal;
vulvar administration; administration onto skin, scar, keloid,
scalp, eye, mouth, nose, vulva, vagina, rectum; and/or
administration into a wound, ulcer, and granulation tissue.
[0187] The compositions may be suitable for administration to hair,
and onto finger or toe nails. Alternatively, the compositions may
be suitable for subcutaneous administration.
Cosmetic Product
[0188] A "cosmetic product," as used herein, include without
limitation, personal care product, skin product, skin cream, skin
gel, skin ointment, skin lotion, anti-aging product, skin
rejuvenation product, skin conditioner, moisturizer, feminine
product, hygiene product, skin patch, skin mask, tissue wipe,
lipstick, mascara, rouge, foundation, blush, eyeliner, lip liner,
lip gloss, lip balm, facial or body powder, sunscreens, sunblocks,
nail polish, mousse, sprays, styling gels, nail conditioner, bath
and shower gels, shampoos, conditioners, cream rinses, hair sprays,
hair dyes and coloring products, soaps, body scrubs, exfoliants,
astringents, depilatories and permanent waving solutions,
antidandruff formulations, anti-sweat and antiperspirant
compositions, shaving, preshaving and after shaving products,
leave-on conditioners, deodorants, cold creams, deodorants,
cleansers, rinses, vulvar product, vaginal product, or the like;
whether in the form of creams, lotions, gels, ointments,
macro-emulsions, micro-emulsions, nano-emulsions, serums, balms,
colloids, solutions, liquids, suspensions, dispersions, compacts,
solids, powders, pencils, spray-on formulations, brush-on
formulations, patches, iontophoretic patches, microprojection
patches, microneedle patches, skin delivery enhancing systems,
bandage, tissue cloths, wipes, masks, aerosols, pastes, soap bars,
cosmetic devices, and/or any other forms readily known to those
skilled in the art.
Medicament
[0189] A "medicament," as used herein, include without limitation
pharmaceutical preparations, carriers for dermatological purposes,
including topical and transdermal application of pharmaceutical
ingredients. These can be in the form of creams, lotions, gels,
ointments, macro-emulsions, micro-emulsions, nano-emulsions,
serums, balms, colloids, solutions, liquids, suspensions,
dispersions, compacts, solids, powders, pencils, spray-on
formulations, brush-on formulations, patches, iontophoretic
patches, microprojection patches, microneedle patches, skin
delivery enhancing systems, bandages, tissue cloths, wipes, masks,
aerosols, pastes, soap bars, medical devices, and/or any other
forms readily known to those skilled in the art.
Suitability for Topical Application
[0190] The term "acceptable substance(s) for topical application",
as used herein, mean that the composition(s) comprising "acceptable
substance(s) for topical application" according to the invention
are suitable for use in contact with human skin and/or human
mucosa; where the skin or the mucosa can be healthy, newborn,
young, old, aged, appear visually different than normal, damaged,
photo-damaged, sunburned, wrinkled, pathologic, diseased, wounded,
atrophic, irritated, compromised, treated with cosmetic product(s),
treated with pharmaceutical product(s), treated with cosmetic
procedure(s), treated with dermatological procedure(s), treated
with a pharmaceutical or medical device(s), surgically treated,
etc. and are absent of significant (consumer-unacceptable) local
intolerabilities to skin or mucosa (i.e., corrosivity, irritation,
allergy), and the like after repeated topical application for
cosmetic, skin care, feminine, or similar uses; or with maximally
low and acceptable (consumer-acceptable) local intolerabilities to
skin or mucosa skin irritation (i.e., irritation, allergy), and the
like after repeated topical application for medical uses of the
composition.
[0191] Local tolerability (e.g., irritation and allergy to skin;
also called contact dermatitis and allergy) in humans can be
determined by acute (1 day) and repetitive (4 to 21 days) patch
testing on the back of humans, and/or during in use tests where the
composition is used as indicated (e.g., for topical use on face,
vulva, vagina, mucosal surface, and/or other body surface areas; or
for wound healing). In case of a medication, safety studies
generally also include animal studies.
[0192] Furthermore, acceptable substance(s) for topical application
means that the compositions comprising "acceptable substance(s) for
topical application" in accordance with the present invention are
without significant physicochemical instability (e.g., significant
changes in color, odor, viscosity, pH, and/or appearance) in the
final packaging (e.g., bottle, tube, pump, jar, airless container,
spray, patch, etc.) during the shelf-life of the product according
to the recommended storage conditions of the product. Significant
physicochemical instability means, that the color, odor, viscosity,
pH, or the appearance changed (increased, decreased) more than 10%
from the time when the composition was prepared and filled into the
final packaging.
[0193] Any of the compositions of the present invention may also
provide good aesthetics and be cosmetically elegant.
[0194] Acceptable substances for topical application or
administration may include suitable excipients and/or carriers
known in the art.
Additional Substances
[0195] The compositions described herein preferably include
Octanoyl Carnosine (or one or more derivatives thereof), optionally
in combination with at least one additional substance suitable for
topical application and/or subcutaneous application. Additional
substance(s) can be inert (e.g., carriers and/or d excipients) or
can be with biological activities (i.e., biologically active agents
and/or active pharmaceutical ingredient). Preferably, the
compositions of the invention may also include additional
biological active agents, including, but not limited to, peptides
other than Octanoyl Carnosine, N-Palmitoyl-GHK, and GEKG (or one or
more derivatives thereof) according to the present invention.
[0196] The terms "substance", "ingredient", "agent" and the like
are used interchangeably herein.
[0197] The compositions of the invention may include one or more
substances, various, conventional or not, which will provide some
benefit to the object of the composition. More specifically, the
combination of Octanoyl Carnosine (or one or more derivatives
thereof) according to the present invention with selected
additional ingredients may lead to an enhanced efficacy as compared
to the use of the Octanoyl Carnosine (or one or more derivatives
thereof) alone. The enhanced efficacy can be additive (the sum of
efficacies of the individual agents alone), or it can be
synergistic (larger than the sum of efficacies of the individual
agents alone). Of course, the decision to include an additional
ingredient and the choice of a specific ingredient depends on the
specific use of the composition and the product formulation and is
well within the routine level of skill in the art.
[0198] In particular examples, the compositions of the present
invention may contain a wide range of additional ingredients. The
2012 International Cosmetic Ingredient Dictionary & Handbook,
14th Edition, as well as the Cosmetic Bench Reference--Directory of
Cosmetic Ingredients (published by Cosmetics & Toiletries)
describes a wide variety of non-limiting cosmetic and
pharmaceutical ingredients commonly used in the skin care, personal
care, feminine care, and dermatology and pharmaceutical industry,
which are available for use in the present invention. Additional
examples can be found in the books provided by the United States
Pharmacopeia (USP), the National Formulary (NF), and other
references for cosmetic and pharmaceutical ingredients known in the
art. Each of these references is herein incorporated by reference
in its entirety. This information is regularly updated by the
addition of new ingredients.
[0199] Exemplary functional classes of such ingredients are, but
are not limited to, abrasive agent, absorbent powder, absorption
base, acidulent, activator, adhesion promoter, agent modulating
cell differentiation, agent modulating cell proliferation, agent
stimulating synthesis of dermal or epidermal macromolecules, agent
preventing degradation of dermal or epidermal macromolecules, agent
acting on microcirculation, agent acting on skin barrier, agent
acting on energy metabolism of cells, agent increasing the
substantivity, antimicrobial sequestering agent, analgesic agent,
anesthetic agent, antacid agent, anti-acne agent, anti-aging agent,
anti-wrinkle agent, anti-atrophy agent, anti-androgen agent,
anti-bacterial agent, anti-scar agent, anti-seborrheic agent,
anti-cracking agent, anti-cellulite agent, anti-stretch mark agent
anti-dandruff agent, anti-foam agent, anti-fungal agent,
anti-histamine agent, anti-inflammatory agent, anti-irritant agent,
anti-microbial agent, anti-mite agents, antibiotic agent, antiviral
agent, antioxidant agent, anti-glycation agent, anti-neoplastic
agent, anti-cancer agent, anti-skin cancer agent, anti-eczema
agent, anti-psoriasis agent, antipollution agent, antiperspirant
agent, anti-pruriginous agent, anti-pruritic agent, antiseptic
agent, antistat agent, astringent, .alpha.-adrenergic receptor
agonist, barrier agent, binding agent, bio-adhesive agents,
botanical agent, botanical extract, biological additive, buffer
agent, bulking agent, calcium sequestering agent, calming agent,
carrier agent, chemical additive, cell lysate, cell culture medium,
conditioned cell culture medium, chelating agent, circulatory
stimulant agent, cleansing agent, collagen stimulating agent,
co-emulsifier agent, colorant, conditioning agent, controlled
release agent, cooling agent, co-solvent, coupling agent, curative
agent, denaturant, deodorant agent, depilatory agent, desquamating
agent, detangler agent, detergent, disinfectant, dispersant, dye
stabilizer, dermatologically acceptable carrier, elastin
stimulating agent, extracellular matrix stimulating agent,
emollient, emulsifier, emulsion stabilizer, enzyme, enzymatic
inhibitor, enzyme-inducing agent, coenzyme, cofactor, essential
oil, exfoliant, fat soluble agent, fiber, film former, fixative,
flavor, foam booster, foam stabilizer, foaming agent, fragrance,
free radicals scavenger, fungicide, gellant, glosser, hair beaching
agent, hair growth promoter, hair colorant, hair conditioning
agent, hair-set polymer, hormone, hormone-like agent, humectant,
hydrophobic agent, hydrotropic agents intermediate agent,
hyaluronic acid stimulating agent, keratolytic agent, lathering
agent, lipolytic agent, lubricant, make-up agent, moisture barrier
agent, moisturizer, muco-adhesive agents, muscle relaxant, natural
moisturizing factor, neutralizer, odor-masking agent, oil, oil
absorbent agent, ointment base, opacifier, organosilicone, oxidant,
oxygen carrier, pearlant agent, perfume, perfume solvent, perfume
stabilizer, peroxide stabilizer, pharmaceutical drug,
photo-sensitizer agent, pigment, pigmenting agent, pearlescent aid,
plant extract, plant derivative, plant tissue extract, plant root
extract, plant seed extract, plant oil, plasticizer, polish agent,
polymer, polymer film former, powder, preservative agent,
propellant, peptide agent, protein agent, reducing agent,
re-fatting agent, regenerator, resin, rosacea inhibitory agent,
scar prevention agent, scalp agent, scrub agent, sabostatic agent,
sequestrant, sex hormone, sex stimulating agent, silicone agent,
silicone replacement agent, skin barrier agent, skin barrier
restoration agent, skin calming agent, skin clarifier, skin
cleanser, skin conditioning agent, skin exfoliating agent, skin
peeling agent, skin healing agent, skin lipid, skin lightening
agent, skin bleaching agent, skin protectant agent, skin purifier
agent, skin smoothing agent, skin calming agent, skin soothing
agent, skin sensate, skin treatment agent, skin penetration
enhancing agent, skin penetration retarding agent, mucosa
penetration enhancing agent, solubilizer, solvent, suspending
agent, sun protection factor booster, soothing agent, spreading
agent, stabilizer, stimulant agent, slimming agent, sunless tanning
agent, sunscreen, sunscreen UVA, sunscreen UVB, broad-band
sunscreen, super-fatting agent, surfactant, amphoteric surfactant,
anionic surfactant, cationic surfactant, non-ionic surfactant,
silicone surfactant, suspending agent, sweetener, tanning
accelerator, thickening agent, thixotrope, tightening agent, toner,
tonic agent, topical delivery system, vasoconstrictor agent, vulvar
soothing agent, vaginal soothing agent, vegetable oil, volatile
agent, viscosity stabilizer, vitamin, vaccine, water proofing
agent, water-soluble agent, water-proofing agent, wax, wetting
agent, whitening agent, wound healing agent, and/or the like.
[0200] Preferably, the additional ingredients should be suitable
for use in contact with human keratinous tissue (hair, nails, skin,
lips, external vulva (mons pubis, labia majora, labia minora))
and/or non-keratinous tissue (vagina, introitus, inner vulva
(vulvar vestibule, clitoris), mouth, anus, etc.), without undue
systemic toxicity local intolerability, and chemical
instability.
[0201] In most instances, the additional substances will include a
cosmetic, dermatologically, and/or pharmaceutically acceptable
carrier either alone or in combination with still other additional
(e.g., inert and/or biologically active) ingredients. The total
amounts of additional ingredients may range from about 90% to about
99.9999%, preferably from about 95% to about 99.999%, and more
preferably from about 99% to about 99.999%, of the composition. In
short, it is the balance of the composition. If carriers (either
singularly, such as water, or complex co-solvents) are used, they
may make up the entire balance of the compositions.
[0202] Non-limiting examples of additional ingredients for some of
the functional classes listed above are provided herein. Additional
examples of additional ingredients can be found in The
International Cosmetic Ingredient Dictionary and Handbook, the
Cosmetic Bench Reference--Directory of Cosmetic Ingredients, the
books provided by the United States Pharmacopeia (USP) and the
National Formulary (NF), and other references for cosmetic and
pharmaceutical ingredients known (and commonly used) in the
art.
[0203] In order to be suitable for use in accordance with the
present invention, the additional ingredients and
carrier/excipients must be further chemically compatible with
Octanoyl Carnosine (or one or more derivatives thereof), with
Palmitoyl-GHK (or one or more derivatives thereof), and/or with
GEKG (SEQ ID NO:1) (or one or more derivatives thereof). Here,
"chemically compatible" means that the additional ingredients do
not lead to a significant chemical degradation (e.g., hydrolysis,
oxidation). For example, a significant chemical degradation would
include more than 10% degradation during the shelf-life period
(e.g., as provided by the expiration date) of the product under the
recommended storage conditions of the product.
[0204] Peptides
[0205] The composition of the present invention can contain
additional peptide(s). Suitable peptides can include, but are not
limited to, di-, tri-, tetra-, penta-, hexa-peptides, and other
oligo- to poly-peptides, and derivatives thereof.
[0206] For example, when included in the present compositions, the
additional peptides are preferably used in amounts ranging from
about 0.000001% to about 10%, more preferably from about 0.000001%
to about 1%, and even more preferably from about 0.00001% to about
0.1% by weight of the composition. The exact content (%) of
peptides to be used in the compositions will depend on the
particular peptide utilized since such agents vary widely in
potency.
[0207] Suitable dipeptides for use herein include, but are not
limited to, carnosine (beta-Ala-His), Tyr-Arg, Val-Trp (see WO
0164178), Asn-Phe, Asp-Phe. Suitable tripeptides for use herein
include, but are not limited to, Arg-Lys-Arg, His-Gly-Gly,
Gly-His-Lys, Gly-Gly-His, Gly-His-Gly, Lys-Phe-Lys. Suitable
tetrapeptides for use herein include, but are not limited to,
Peptide E, Arg-Ser-Arg-Lys (SEQ ID NO:10), Gly-Gln-Pro-Arg (SEQ ID
NO:9). Suitable pentapeptides include, but are not limited to,
Lys-Thr-Thr-Lys-Ser (SEQ ID NO:11). Suitable hexapeptides include,
but are not limited to, Val-Gly-Val-Ala-Pro-Gly (SEQ ID NO:12) and
such as those disclosed in FR 2854897 and US 2004/0120918.
[0208] Other suitable peptides for use herein include, but are not
limited to, lipophilic derivatives of peptides, preferably
octanoyl, decanoyl, lauroyl, myristoyl and palmitoyl derivatives,
and metal complexes of the aforementioned (e.g., copper complex of
the tripeptide His-Gly-Gly). Preferred dipeptide derivatives
include N-Palmitoyl-beta-Ala-His, N-Acetyl-beta-Ala-His,
N-Acetyl-Tyr-Arg-hexadecylester (WO 9807744, U.S. Pat. No.
6,372,717). Preferred tripeptide derivatives include the copper
derivative of His-Gly-Gly, N-Elaidoyl-Lys-Phe-Lys and its analogs
of conservative substitution, N-Acetyl-Arg-Lys-Arg-NH2,
N-Biot-Gly-His-Lys (WO0058347) and derivatives thereof. Suitable
tetrapeptide derivatives for use herein include, but are not
limited to N-palmitoyl-Gly-Gln-Pro-Arg, suitable pentapeptide
derivatives for use herein include, but are not limited to
N-Palmitoyl-Lys-Thr-Thr-Lys-Ser (WO 0015188 and U.S. Pat. No.
6,620,419), N-Palmitoyl-Tyr-Gly-Gly-Phe-X with X Met or Leu (SEQ ID
NO:13), or mixtures thereof. Suitable hexapeptide derivatives for
use herein include, but are not limited to
N-Palmitoyl-Val-Gly-Val-Ala-Pro-Gly and derivatives thereof.
[0209] The peptides can be obtained from any supplier of
commercially available cosmetic and pharmaceutical peptides,
peptide mixtures or derivatives thereof; including but not limited
to Atrium, Unipex, Lucas Meyer Cosmetics, Biotechnologies, Sederma,
Croda, Grant Industries, Pentapharm, DSM, Evonik, Lipotec, Symrise,
BASF, ISP, Helix BioMedix, Oriflame, Seppic, Solabia, Procyte, EMD
Chemicals, Corium Peptides, etc.; or can be directly obtained by
custom synthesis. When using commercially available cosmetic and
pharmaceutical peptides, the preferred composition generally
contains the additional peptide(s) in the concentration range as
recommended by the peptide supplier.
[0210] Additional examples of suitable peptides can be also found
in the chapter by F. Gorohhui and H. I. Maibach in the Textbook of
Aging (2010, Springer), in Clinics in Dermatology 2009, 27,
485-495, or numerous other scientific articles, communications,
patent applications, granted patents on peptides for cosmetic or
medical uses (incorporated herein by reference).
[0211] Ascorbates and Other Vitamins
[0212] The compositions of the present invention may contain one or
more vitamins, such as ascorbates (e.g., vitamin C, vitamin C
derivatives, ascorbic acid, ascorbyl glucoside, ascorbyl palmitate,
magnesium ascorbyl phosphate, sodium ascorbyl phosphate,
tetrahexadecyl ascorbate, ascorbyl 3-aminopropyl phosphate),
vitamin B, vitamin B derivatives, vitamin B1 to vitamin B12 and
theirs derivatives, vitamin K, vitamin K derivatives, vitamin H,
vitamin D, vitamin D3, vitamin D derivatives, vitamin E, vitamin E
derivatives, and pro-vitamins thereof, such as panthenol and
mixtures thereof. The vitamin compounds may be included as the
substantially pure material, or as an extract obtained by suitable
physical and/or chemical isolation from natural (e.g., plant)
sources. In one example, when vitamin compounds are present in the
compositions of the instant invention, the compositions contain
from about 0.0001% to about 25%, more preferably from about 0.001%
to about 10%, still more preferably from about 0.01% to about 5%,
and still more preferably from about 0.1% to about 1%, by weight of
the composition, of the vitamin compound. The exact content (%) of
ascorbates and other vitamins to be used in the compositions will
depend on the particular ascorbate and vitamin utilized since such
agents vary widely in potency.
[0213] Sunscreen Actives
[0214] The compositions of the subject invention may optionally
contain a sunscreen active. As used herein, "sunscreen active"
includes both sunscreen agents and physical sunblocks. Suitable
sunscreen actives may be organic or inorganic. A wide variety of
conventional organic or inorganic sunscreen actives are suitable
for use herein. In one example, the composition contains from about
0.1% to about 25%, more typically from about 0.5% to about 10% by
weight of the composition, of the sunscreen active. Exact amounts
will vary depending upon the sunscreen chosen and the desired Sun
Protection Factor (SPF). The organic UV-screening agents which are
more particularly preferred are chosen from the following
compounds: ethylhexyl salicylate, butyl methoxydibenzoylmethane,
ethylhexyl methoxycinnamate, octocrylene, phenylbenzimidazole
sulphonic acid, terephthalylidene dicamphor sulphonic,
benzophenone-3, benzophenone-4, benzophenone-5,4-methylbenzylidene
camphor, benzimidazilate, anisotriazine, Ethylhexyl triazone,
diethylhexyl butamido triazone, methylene bis-benzotriazolyl
tetramethylbutylphenol, drometrizole trisiloxane, and mixtures
thereof.
[0215] The inorganic sunscreen agents which may be used in the
composition according to the invention are in particular
nanopigments (mean size of the primary particles: generally between
5 nm and 100 nm, preferably between 10 nm and 50 nm; or their
aggregates) of coated or uncoated metal oxides such as for example
nanopigments of titanium oxide (amorphous or crystallized in the
form of rutile and/or anatase), iron, zinc, zirconium or cerium
oxides and mixtures thereof. Coating agents are moreover alumina
and/or aluminum stearate, and silicones.
[0216] Anti-Wrinkle Actives and Anti-Atrophy Actives
[0217] The compositions of the present invention can contain a one
or more anti-wrinkle actives or anti-atrophy actives. Exemplary
anti-wrinkle/anti-atrophy actives suitable for use in the
compositions of the present invention include amino acids, N-acetyl
derivatives of amino acids (e.g., N-acetyl-cysteine), hydroxy acids
(e.g., alpha-hydroxy acids such as lactic acid and glycolic acid or
beta-hydroxy acids such as salicylic acid and salicylic acid
derivatives such as the octanoyl derivative, lactobionic acid),
keto acids (e.g., pyruvic acid), phytic acid, ascorbic acid
(vitamin C), retinoids (e.g., retinoic acid, tretinoin,
isotretinoin, adapalene, retinol, retinylaldehyde,
retinylpalmitate, and other retinoid derivatives), kinetin
(N6-furfuryladenine), zeatin and their derivatives (e.g.,
furfurylamino-tetrahydropyranyladenine), niacinamide
(nicotinamide); growth factors and cytokines (e.g., TGF-beta 1, 2
and 3, EGF, FGF-2, PDGF, IL-1, IL-6, IL-8, IGF-1, IGF-2, etc.),
cell lysates (e.g., dermal fibroblast cell lysate, stem cell
lysate, processed skin cell proteins (PSP.RTM.), etc.), conditioned
cell culture mediums (e.g., conditioned cell culture medium from
dermal fibroblasts, conditioned cell culture medium from stem cells
(e.g., epidermal stem cells, adipose stem cells, mesenchymal stem
cells, etc.); cosmetic ingredients marketed under the trade names
Nouricel-MD.RTM., TNS.RTM., or CCM.TM. Complex; etc.); cell
extracts, stem cell extracts, components from stem cells;
ingredients stimulating epidermal or other human adult stem cells;
skin conditioning agents, stilbenes, cinnamates, ingredients
activating sirtuin 1 (e.g., resveratrol); ingredients improving the
functioning of the mitochondria; dimethylaminoethanol, synthetic
anti-aging peptides, peptides from natural sources (e.g., soy
peptides), and salts of sugar acids (e.g., Mn gluconate, Zn
gluconate), lipoic acid; lysophosphatidic acid, vitamin B3
compounds, and other vitamin B compounds (e.g., thiamine (vitamin
B1), pantothenic acid (vitamin B5), riboflavin (vitamin B2), and
their derivatives and salts (e.g., HCl salts or calcium salts).
[0218] When anti-wrinkle/anti-atrophy compounds are present in the
compositions of the instant invention, the compositions comprise
from about 0.0001% to about 25%, more preferably from about 0.001%
to about 10%, still more preferably from about 0.01% to about 5%,
and still more preferably from about 0.1% to about 1%, by weight of
the composition, of the anti-wrinkle/anti-atrophy compound. The
exact content (%) of anti-wrinkle/anti-atrophy agents to be used in
the compositions will depend on the particular
anti-wrinkle/anti-atrophy agent utilized since such agents vary
widely in potency.
[0219] Humectants, Moisturizers, and Conditioning Agents
[0220] The compositions of the present invention can contain a safe
and effective amount of a conditioning agent selected from, for
example, humectants, moisturizers, and skin conditioners. A variety
of these materials can be employed and can be present at a level of
from about 0.01% to about 80%, more preferably from about 0.1% to
about 25%, and still more preferably from about 0.5% to about 10%,
by weight of the composition. The exact content (%) of humectants,
moisturizers, and conditioning agents to be used in the
compositions will depend on the humectant, moisturizer, and
conditioning agent utilized since such agents vary widely in
potency.
[0221] Humectants are ingredients that help maintain moisture
levels in skin. Humectants can be selected from the group
consisting of polyhydric alcohols, water soluble alkoxylated
nonionic polymers, and mixtures thereof. Polyhydric alcohols useful
herein include polyhdroxy alcohols aforementioned and glycerin,
hexylene glycol, ethoxylated glucose, 1,2-hexane diol, dipropylene
glycol, trehalose, diglycerin, maltitol, maltose, glucose,
fructose, sodium chondroitin sulfate, sodium hyaluronate, sodium
adenosine phosphate, sodium lactate, pyrrolidone carbonate,
glucosamine, cyclodextrin, and mixtures thereof. Water soluble
alkoxylated nonionic polymers useful herein include polyethylene
glycols and polypropylene glycols having a molecular weight of up
to about 1000 such as those with CTFA names PEG-200, PEG-400,
PEG-600, PEG-1000, and mixtures thereof. Additional humectants
include acetyl arginine, algae extract, aloe barbadensis leaf
extract, 2,3-butanediol, chitosan lauroyl glycinate, diglycereth-7
malate, diglycerin, diglycol guanidine succinate, erythritol,
fructose, glucose, glycerin, honey, hydrolyzed proteins,
hydroxypropyltrimonium hyaluronate, inositol, lactitol, maltitol,
maltose, mannitol, mannose, methoxy polyethylene glycol,
myristamidobutyl guanidine acetate, polyglyceryl sorbitol,
potassium pyrollidone carboxylic acid (PCA), propylene glycol,
butylene glycol, sodium pyrollidone carboxylic acid (PCA),
sorbitol, sucrose, dextran sulfate (i.e., of any molecular weight),
natural moisturizing factors, and/or urea.
[0222] Skin conditioners can include, but are not limited to,
guanidine, urea, glycolic acid, glycolate salts (e.g., ammonium and
quaternary alkyl ammonium), salicylic acid, lactic acid, lactate
salts (e.g., ammonium and quaternary alkyl ammonium), aloe vera in
any of its variety of forms (e.g., aloe vera gel), polyhydroxy
alcohols such as sorbitol, mannitol, xylitol, erythritol,
hexanetriol, butanetriol, propylene glycol, butylene glycol,
hexylene glycol and the like, polyethylene glycols, propoxylated
glycerols, sugars (e.g., melibiose), starches, sugar and starch
derivatives (e.g., alkoxylated glucose, fructose, glucosamine),
C1-C30 monoesters and polyesters of sugars and related materials,
hyaluronic acid, lactamide monoethanolamine, acetamide
monoethanolamine, panthenol, dexpanthenol, allantoin, and mixtures
thereof. Skin conditioners can also include fatty acids, fatty acid
esters, lipids, ceramides, cholesterol, cholesterol esters, bee
wax, petrolatum, and mineral oil.
[0223] Emollients
[0224] One or more emollients may also be included in the topical
compositions described herein. An emollient generally refers to an
ingredient that can help skin maintain a soft, smooth, and pliable
appearance. Emollients typically remain on the skin surface, or in
the stratum corneum, and act as a moisturizer, or lubricant and
reduce flaking. Some examples of emollients include acetyl
arginine, acetylated lanolin, algae extract, apricot kernel oil
polyethylene glycol-6 esters, avocado oil polyethylene glycol-11
esters, bis-polyethylene glycol-4 dimethicone, butoxyethyl
stearate, glycol esters, alkyl lactates, caprylyl glycol, cetyl
esters, cetyl laurate, coconut oil polyethylene glycol-10 esters,
alkyl tartrates, diethyl sebacate, dihydrocholesteryl butyrate,
dimethiconol, dimyristyl tartrate, disteareth-5 lauroyl glutamate,
ethyl avocadate, ethylhexyl myristate, glyceryl isostearates,
glyceryl oleate, hexyldecyl stearate, hexyl isostearate,
hydrogenated palm glycerides, hydrogenated soy glycerides,
hydrogenated tallow glycerides, isostearyl neopentanoate,
isostearyl palmitate, isotridecyl isononanoate, laureth-2 acetate,
lauryl polyglyceryl-6 cetearyl glycol ether, methyl gluceth-20
benzoate, mineral oil, palm oil, coconut oil, myreth-3 palmitate,
octyldecanol, octyldodecanol, odontella aurita oil, 2-oleamido-1,3
octadecanediol, palm glycerides, polyethylene glycol avocado
glycerides, polyethylene glycol castor oil, polyethylene
glycol-22/dodecyl glycol copolymer, polyethylene glycol shea butter
glycerides, phytol, raffinose, stearyl citrate, sunflower seed oil
glycerides, petrolatum, silicon oils including but not limited to
caprylyl methicone, and/or tocopheryl glucoside.
[0225] Anti-Oxidants, and Radical Scavengers
[0226] The compositions of the present invention may include an
anti-oxidant/radical scavenger. In one example, the composition
contains from about 0.001% to about 25%, more preferably from about
0.01% to about 10%, and still more preferably from about 0.1% to
about 5%, of an anti-oxidant/radical scavenger. The exact content
(%) of anti-oxidant/radical scavengers to be used in the
compositions will depend on the particular anti-oxidant/radical
scavenger utilized since such agents vary widely in potency.
[0227] Anti-oxidants/radical scavengers may include but are not
limited to ascorbic acid (vitamin C) and its salts, ascorbyl esters
of fatty acids, and other ascorbic acid derivatives (e.g.,
magnesium ascorbyl phosphate, sodium ascorbyl phosphate, ascorbyl
sorbate, ascorbyl palmitate, tetrahexyldecyl ascorbate, etc.),
tocopherol (vitamin E), tocopherol sorbate, tocopherol acetate,
other esters of tocopherol, beta-carotene, butylated hydroxy
benzoic acids and their salts, ferulic acid, peroxides including
hydrogen peroxide, perborate, thioglycolates, persulfate salts,
6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid
(commercially available under the trade name Trolox.TM.), gallic
acid and its alkyl esters, especially propyl gallate, uric acid and
its salts and alkyl esters, amines (e.g., N,N-diethylhydroxylamine,
amino-guanidine), nordihydroguaiaretic acid, bioflavonoids,
sulfhydryl compounds (e.g., glutathione), dihydroxy fumaric acid
and its salts, lysine pidolate, arginine pilolate, amino acids,
silymarin, lysine, 1-methionine, proline, superoxide dismutase,
sorbic acids and its salts, lipoic acid, olive extracts, tea
extracts, resveratrol, polyphenols such as proanthocyanidine from
pine bark, carotenoids, curcumin compounds such as
tetrahydrocurcumin, coenzyme Q10, OCTA (L-2-oxo-4-thiazolidine
carboxylic acid), selenium, creatine, glutathione, N-acetyl
cysteine, N-acetyl cysteine esters, dimethylmethoxy chromanol,
lipoic acid, melanin; plant extracts containing polyphenols
including but not limited to coffee berry extracts, green tea
extracts, rosemary extracts, witch hazel extracts, and grape
skin/seed extracts, may be used. Preferred anti-oxidants/radical
scavengers can be selected from esters of ascorbic acid,
tocopherol, ferulic acid, polyphenols, creatine, and their
derivatives; as well as plant extracts containing polyphenols such
as green tea extract.
[0228] Antimicrobial Peptide Sequestering Agents
[0229] Antimicrobial peptide sequestering compounds may include but
are not limited to a sulfated or polysulfated monosaccharide, and
salts and complexes thereof; a sulfated or polysulfated
disaccharide, and salts and complexes thereof; a sulfated or
polysulfated polysaccharide, and salts and complexes thereof; a
dextran sulfate (e.g., sodium dextran sulfate), and salts and
complexes thereof; chondroitin sulfate, and salts and complexes
thereof; pentosan polysulfate, and salts and complexes thereof;
sucrose sulfate (e.g., any sucrose sulfate such as sucrose
octasulphate other than aluminum sucrose sulfate), and salts and
complexes thereof; a fucoidan, and salts and complexes thereof; a
sulfated galactan, and salts and complexes thereof; a carrageenans
(e.g., Chondrus Crispus), and salts and complexes thereof; starch
sulfate, and salts and complexes thereof; cellulose sulfate, and
salts and complexes thereof; a sulfated glycosaminoglycan, and
salts and complexes thereof; a heparin; a heparan sulfate; sulfated
glucan; or any combinations thereof. The antimicrobial peptide
sequestering compound may include a plant extract, an algae
extract, an aloe vera (barbadensis) extract, a cactus extract, or a
shark or fish cartilage extract. The antimicrobial peptide
sequestering compound may also be a sulfated or polysulfated
polymer (e.g., poly(vinyl sulfate), poly(anethole sulfonate)).
Suitable polymeric sulfonic acid that can be used in the methods
and compositions described herein are hydrophobically modified
polymeric sulfonic acids such as Aristoflex.RTM. HMP or
Aristoflex.RTM. AVC (Clamant). Alternatively, the antimicrobial
peptide sequestering compound is a phosphate or polyphosphate
(e.g., a monosaccharide phosphate, a disaccharide phosphate, a
polysaccharide phosphate, a glycerophosphate salt, or a starch
phosphate). Suitable examples of starch phosphates include, but are
not limited to hydroxypropyl starch phosphates (i.e., Structure XL
(National Starch, LCC)). The antimicrobial peptide sequestering
compound may also be a phospholipid such as phosphatidylcholine or
lecithin. Further, the antimicrobial peptide sequestering compound
can be a carboxylate, a polyhydroxy acid, hyaluronic acid,
alginate, and/or polylactic acid. Most preferably, the
antimicrobial peptide sequestering compounds are between 100 to
10,000 g per mol. Sodium dextran sulfate of about 5000 to 10'000 g
per mol is one of the most preferred antimicrobial peptide
sequestering compound.
[0230] Rosacea Inhibitory Agents, and .alpha.-Adrenergic Receptor
Agonists
[0231] Rosacea inhibitory agents, include but are not limited to,
metronidazole, sulfacetamide, sodium sulfacetamide, sulfur, dapson,
doxycycline, minocycline, clindamycin, clindamycin phosphate,
erythromycin, tetracylclines, azelaic acid, calcium dobesilate,
maleic acid, and any compatible combinations thereof);
.alpha.-adrenergic receptor agonists (e.g., clonidine, amphetamine,
doxtroamphetamine, apraclonidine, dipivefrin, .alpha.-methyldopa,
oxymetazoline, oxymetazoline hydrochloride, methoxamine,
metaraminol, medetomidine, dexmedetomidine, ethylnorepinephrine,
guanfacine, guanabenz, phenylephrine, phenylephrine hydrochloride,
ephedrine, epinine, epinephrine, ethylnorepinephrine, levarterenol,
lofexidine, norepinephrine, norphenylephrine, norephedrine,
phenylpropanolamine, pemoline, propylhexadrine, pseudoephedrine,
methamphetamine, .alpha.-methylnorepinephrine, methylphenidate,
mephentermine, midodrine, mivazerol, moxonidine, desglymidodrine,
tetrahydrozoline, tetrahydrozoline hydrochloride, cirazoline,
amidephrine, brimonidine, brimonidine tartrate, naphazoline,
isoproterenol, xylazine, xylometazoline, and/or tizanidine);
chemicals and botanical extracts with vasoconstrictor properties
including, but not limited to, corticosteroids, ephedrine,
pseudoephedrine, caffeine, and/or escin; ephedra, phedra sinica,
hamamelis viginiana, hydrastis canadensis, lycopus virginicus,
aspidosperma quebracho, cytisus scoparius, raphanus sativus linn
(radish leave extracts), horse chestnut extracts, etc., as well as
any compatible combinations thereof; and/or a nasal and/or sinus
decongestant.
[0232] Skin Lightening Agents, and Skin Bleaching Agents
[0233] The compositions of the present invention may contain a skin
lightening agent. Suitable skin lightening agents include, but are
not limited to, ascorbic acid and derivatives thereof; kojic acid
and derivatives thereof; resorcinol and derivatives thereof
(including but not limited to 4-ethyl resorcinol, 4-butyl
resorcinol, 4-hexyl resorcinol, 4-octyl resorcinol, 4-decyl
resorcinol, 6-methyl resorcinol, 6-ethyl resorcinol, 6-butyl
resorcinol, 6-hexyl resorcinol, 6-octyl resorcinol, 6-decyl
resorcinol, 4-phenylethyl resorcinol), retinoic acid and
derivatives thereof (e.g., retinol, retinyl palmitate), L-leucine
and derivatives thereof (e.g., N-acyl derivatives of L-leucine,
esters of L-leucine, etc.), glycine and derivatives thereof,
disodium glycerophosphate and derivatives thereof, undecenoyl
phenylalanine, arbutin and derivatives thereof (e.g.,
dehydroxyarbutin), niacinamide and derivatives thereof,
hydroquinone; mequinol, glabridin, aleosin, curcumin, genistein,
ethyl linoleate, tranexaminic acid, azelaic acid, resveratrol and
derivatives thereof (e.g., oxyresveratrol), N-acetyl glucosamine,
4-isopropylcetchol, 4-ethoxybenzaldehyde, 2-ethoxybenzaldehyde,
4-propoxybenzaldehyde, alpha-hydroxyacids (e.g., glycolic acid,
lactic acid, etc.), salicylic acid, polyphenols; and/or various
plant extracts, such as those from licorice, grape seed, mulberry,
soy, green tea, and/or bear berry; and/or any ingredient or
combination thereof.
[0234] When used, the compositions preferably contain from about
0.01% to about 15%, more preferably from about 0.1% to about 10%,
also preferably from about 0.5% to about 5%, by weight of the
composition, of a skin lightening agent. The exact content (%) of
skin lightening agents to be used in the compositions will depend
on the particular skin lightening agent utilized since such agents
vary widely in potency.
[0235] Skin Protectants
[0236] Suitable skin protectant agents for use in the compositions
described herein include, for example, a compound that protects
injured or exposed skin or mucous membrane surfaces from harmful or
irritating external compounds. Representative examples include
algae extract, allantoin, camellia sinensis leaf extract,
cerebrosides, dimethicone, glucuronolactone, glycerin, kaolin,
lanolin, malt extract, mineral oil, petrolatum, white petrolatum,
potassium gluconate, colloidal oat meal, calamine, cocoa butter,
starch, zinc oxide, zinc carbonate, zinc acetate, and/or talc.
[0237] Desquamation Actives, Keratolytic Agents, and Peeling
Agents
[0238] A desquamating/keratolytic active may be added to the
compositions of the present invention. In one example, the
composition contains from about 0.01% to about 30%, preferably from
about 0.1% to about 10%, more preferably from about 0.5% to about
5%, by weight of the composition, of a desquamating/keratolytic
active. The exact content (%) of desquamating/keratolytic agents to
be used in the compositions will depend on the particular
desquamating/keratolytic agent utilized since such agents vary
widely in potency.
[0239] Examples of useful keratolytic and/or desquamating agents
include urea, salicylic acid and alkyl derivatives thereof,
saturated and unsaturated monocarboxylic acids, saturated and
unsaturated bicarboxylic acids, tricarboxylic acids, alpha
hydroxyacids and beta hydroxyacids of monocarboxylic acids, alpha
hydroxyacids and beta hydroxyacids of bicarboxylic acids, alpha
hydroxyacids and beta hydroxyacids of tricarboxylic acids,
ketoacids, alpha ketoacids, beta ketoacids, of the polycarboxylic
acids, of the polyhydroxy monocarboxylic acids, of the polyhydroxy
bicarboxylic acids, of the polyhydroxy tricarboxylic acids.
Resorcinol and its low-molecular weight derivatives are other
examples of useful keratolytic and/or desquamating agents.
[0240] Preferred keratolytic agents are selected from the group
containing glycolic acid, tartaric acid, salicylic acid, citric
acid, lactic acid, pyruvic acid, gluconic acid, glucuronic acid,
malic acid, mandelic acid, oxalic acid, malonic acid, succinic
acid, acetic acid, phenol, resorcinol, retinoic acid, adapalene,
trichloroacetic acid, 5-fluoro uracil, azelaic acid. Keratolytic
agents are also the salts, esters, possible cis- or trans-forms,
racemic mixtures and/or the relative dextrorotatory or levorotatory
forms of the above listed compounds. Such substances can be used
singularly or in associations with each other.
[0241] Anti-Inflammatory Agents
[0242] An anti-inflammatory agent may be added to the compositions
of the present invention. In one example, an anti-inflammatory
agent is added at a level of from about 0.01% to about 10%,
preferably from about 0.5% to about 5%, by weight of the
composition. The exact content (%) of anti-inflammatory agents to
be used in the compositions will depend on the particular
anti-inflammatory agent utilized since such agents vary widely in
potency.
[0243] Steroidal anti-inflammatory agents can include, but are not
limited to, corticosteroids such as hydrocortisone,
hydroxyltriamcinolone, alpha-methyl dexamethasone,
dexamethasone-phosphate, beclomethasone dipropionates, clobetasol
valerate, desonide, desoxymethasone, desoxycorticosterone acetate,
dexamethasone, dichlorisone, diflorasone diacetate, diflucortolone
valerate, fluadrenolone, fluclorolone acetonide, fludrocortisone,
flumethasone pivalate, fluosinolone acetonide, fluocinonide,
flucortine butylesters, fluocortolone, fluprednidene
(fluprednylidene) acetate, fluradrenolone, halcinonide,
hydrocortisone acetate, hydrocortisone butyrate,
methylprednisolone, triamcinolone acetonide, cortisone,
cortodoxone, flucetonide, fludrocortisone, difluorosone diacetate,
fluradrenolone, fludrocortisone, diflurosone diacetate,
fluradrenolone acetonide, medrysone, amcinafel, amcinafide,
betamethasone and the balance of its esters, chloroprednisone,
chlorprednisone acetate, clocortelone, clescinolone, dichlorisone,
diflurprednate, flucloronide, flunisolide, fluoromethalone,
fluperolone, fluprednisolone, hydrocortisone valerate,
hydrocortisone cyclopentylpropionate, hydrocortamate, meprednisone,
paramethasone, prednisolone, prednisone, beclomethasone
dipropionate, triamcinolone, and mixtures thereof may be used. One
of the preferred steroidal anti-inflammatory for use is
hydrocortisone.
[0244] In addition, non-steroidal anti-inflammatory agents can be
useful herein. The varieties of compounds encompassed by this group
are well known to those skilled in the art. Specific non-steroidal
anti-inflammatory agents that can be useful in the composition of
the present invention include, but are not limited to, diclofenac,
indomethacin, oxicams such as piroxicam, salicylates such as
aspirin; acetic acid derivatives such as felbinac, fenamates such
as etofenamate, flufenamic acid, mefenamic acid, meclofenamic acid,
tolfenamic acid; propionic acid derivatives such as ibuprofen,
naproxen, pyrazoles, and mixtures thereof. Mixtures of these
non-steroidal anti-inflammatory agents may also be employed, as
well as the dermatologically acceptable salts and esters of these
agents. For detailed disclosure of the chemical structure,
synthesis, side effects, etc. of non-steroidal anti-inflammatory
agents, one may refer to standard texts, including
Anti-inflammatory and Anti-Rheumatic Drugs, K. D. Rainsford, Vol.
I-III, CRC Press, Boca Raton, (1985), and Anti-inflammatory Agents,
Chemistry and Pharmacology, 1, R. A. Scherrer, et al., Academic
Press, New York (1974).
[0245] Finally, so-called "natural" anti-inflammatory agents are
useful in methods of the present invention. Such agents may
suitably be obtained as an extract by suitable physical and/or
chemical isolation from natural sources (e.g., plants, fungi,
by-products of microorganisms) or can be synthetically prepared.
For example, candelilla wax, bisabolol (e.g., alpha bisabolol),
aloe vera, plant sterols (e.g., phytosterol), kola extract,
chamomile, red clover extract, sea whip extract, licorice extract,
and tea extract may be used.
[0246] Anti-inflammatory agents useful herein include allantoin and
compounds of the Licorice, including glycyrrhetic acid,
glycyrrhizic acid, and derivatives thereof (e.g., salts and
suitable esters). Additional anti-inflammatory agents include
diosgenol, saponines, sapogenines, lignanes, triterpenes
saponosides and genines.
[0247] Additional examples of anti-inflammatory agents can include
anti-inflammatory interleukins (e.g., IL-lra, IL-10);
anti-inflammatory fatty acids (e.g., linoleic acid, linolenic acid)
and their derivatives (e.g., esters), isoprenylcystein analogues
(i.e., N-acetyl-S-farnesyl-L-cysteine), aromatic aldehydes with
anti-inflammatory properties (e.g., 4-methoxy benzaldehyde,
4-ethoxy benzaldehyde, 4-butoxy benzaldehyde, 4-penthoxy
benzaldehyde), as well as any compatible combinations thereof.
[0248] Anti-Acne Actives
[0249] The compositions of the present invention can contain one or
more anti-acne actives. Examples of useful anti-acne actives
include resorcinol, sulfur, erythromycin, salicylic acid, benzoyl
peroxide, retinoic acid, tretinoin, alpha-hydroxy acids (e.g.,
glycolic acid, lactic acid), dehydroacetic acid and zinc. When
anti-acne compounds are present in the compositions of the instant
invention, the compositions contain from about 0.0001% to about
50%, more preferably from about 0.001% to about 20%, still more
preferably from about 0.01% to about 10%, and still more preferably
from about 0.1% to about 5%, by weight of the composition, of the
anti-acne compound. The exact content (%) of anti-acne actives to
be used in the compositions will depend on the particular
antimicrobial, anti-bacterial and anti-acne active utilized since
such agents vary widely in potency.
[0250] Antimicrobial, Anti-Bacterial and Anti Fungal Actives
[0251] The compositions of the present invention can contain one or
more anti-fungal or anti-microbial actives. A safe and effective
amount of an antimicrobial or antifungal active can be added to the
present compositions. For example, the composition contains from
about 0.001% to about 10%, preferably from about 0.01% to about 5%,
and more preferably from about 0.05% to about 2%, by weight of the
composition, of an antimicrobial or antifungal active. The exact
content (%) of antimicrobial, anti-bacterial and anti-fungal
actives to be used in the compositions will depend on the
particular antimicrobial, anti-bacterial and anti-fungal active
utilized since such agents vary widely in potency.
[0252] Suitable anti-microbial actives include, but are not limited
coal to tar, sulfur, aluminum chloride, gentian violet, octopirox
(piroctone olamine), 3,4,4'-trichlorocarbanilide (trichlosan),
triclocarban, ciclopirox olamine, undecylenic acid and it's metal
salts, potassium permanganate, selenium sulphide, sodium
thiosulfate, propylene glycol, oil of bitter orange, urea
preparations, griseofulvin, 8-hydroxyquinoline ciloquinol,
thiobendazole, thiocarbamates, haloprogin, polyenes,
hydroxypyridone, morpholine, benzylamine, allylamines (such as
terbinafine), tea tree oil, clove leaf oil, coriander, palmarosa,
berberine, thyme red, cinnamon oil, cinnamic aldehyde, citronellic
acid, hinokitol, ichthyol pale, iodopropynyl butylcarbamate,
azelaic acid, isothiazalinones such as octyl isothiazolinone and
azoles, parabens (e.g., methylparaben, ethylparaben, etc.), glycols
(e.g., hexylenglycol, ethylhexylglycerin), and combinations
thereof.
[0253] For example, suitable agents with anti-fungal properties are
ketoconazole, naftifine hydrochloride, oxiconazole nitrate,
sulconazole nitrate, urea, terbinafine hydrochloride, selenium
sulfide. Suitable agents with anti-mite properties are crotamiton,
ivermectin, and permethrin.
[0254] One or more anti-fungal or anti-microbial active is combined
with an anti-dandruff active selected from polyvalent metal salts
of pyrithione.
[0255] Anesthetics
[0256] The compositions of the present invention may also contain a
safe and effective amount of a topical anesthetic. Examples of
topical anesthetic drugs include benzocaine, lidocaine,
bupivacaine, chlorprocaine, dibucaine, etidocaine, mepivacaine,
tetracaine, dyclonine, hexylcaine, procaine, cocaine, ketamine,
pramoxine, phenol; and pharmaceutically acceptable salts thereof;
benzyl alcohol, camphor, menthol, resorcinol; and appropriate
combinations thereof.
[0257] Plant Extracts and Vegetable Extracts
[0258] The compositions of the present invention may also contain a
safe amount of a plant extract and vegetable extract. Examples of
plant or vegetable extracts include extracts obtained from ivy (in
particular English Ivy (Hedera Helix)), Chinese thorowax (Bupleurum
chinensis), barley, Bupleurum Falcatum, arnica (Arnica Montana L),
rosemary (Rosmarinus officinalis N), marigold (Calendula
officinalis), sage (Salvia officinalis L), soy, ginseng (Panax
ginseng), ginko biloba, St.-John's-Wort (Hyperycum Perforatum),
butcher's-broom (Ruscus aculeatus L), European meadowsweet
(Filipendula ulmaria L), big-flowered Jarva tea (Orthosiphon
Stamincus Benth), algae (Fucus Vesiculosus), birch (Betula alba),
green tea, white tea, fermented tea, cola nuts (Cola Nipida),
horse-chestnut, bamboo, spadeleaf (Centella asiatica), heather,
fucus, willow, witch hazel, wild yam, mouse-ear, escine, cangzhu,
chrysanthellum indicum, plants of the Armeniacea genus,
Atractylodis Platicodon, Sinnomenum, Pharbitidis, Flemingia, Coleus
such as C. Forskohlii, C. blumei, C. esquirolii, C. scutellaroides,
C. xanthantus and C, Barbatus, root of Coleus barbatus, Ballote,
Guioa, Davallia, Terminalia, Barringtonia, Trema, antirobia,
cecropia, argania, dioscoreae such as Dioscorea opposita or
Mexican, Ammi visnaga, Centella asiatica and Siegesbeckia, in
particular Siegesbeckia orientalis, the family of Ericaceae in
particular bilberry extracts (Vaccinium angustifollium) or
Arctostaphylos uva ursi, aloe vera, plant sterols (e.g.,
phytosterol), Manjistha (extracted from plants in the genus Rubia,
particularly Rubia Cordifolia), and Guggal (extracted from plants
in the genus Commiphora, particularly Commiphora Mukul), kola
extract, chamomile, red clover extract, Piper methysticum, Bacopa
monieri extract, sea whip, Glycyrrhiza glabra, mulberry, melaleuca
(tea tree), mushroom extracts, Larrea divaricata, Rabdosia
rubescens, euglena gracilis, Fibraurea recisa Hirudinea, Chaparral
Sorghum, sun flower extract, Enantia chlorantha, Mitracarpe of
Spermacocea genus, Buchu barosma, Lawsonia inermis L., Adiantium
Capillus-Veneris L., Chelidonium majus, Luffa cylindrical, Japanese
Mandarin (Citrus reticulata Blanco var. unshiu), broccoli extract,
Camelia sinensis, Imperata cylindrical, Glaucium Flavum, Cupressus
Sempervirens, Polygonatum multiflorum, loveyly hemsleya, Sambucus
Nigra, Phaseolus lunatus, Centaurium, Macrocystis Pyrifera, Turnera
Diffusa, Anemarrhena asphodeloides, Portulaca pilosa, Humulus
lupulus, Coffee Arabica, coffee berry, black berry, Ilex
Paraguariensis; and so on.
[0259] Oils and Lipids
[0260] The oil phase can contain any cosmetic or dermatological oil
or a mixture thereof. Examples of such oils include but are not
limited to aliphatic hydrocarbons such as liquid paraffin,
squalene, squalane, vaseline and ceresin; silicon oils such as
dimethicone and cyclomethicones; vegetable oils such as avocado
oil, apricot oil, almond oil, borage oil, borage seed oil, camellia
oil, canola oil, castor oil, coconut oil, cocoa butter, corn oil,
cottonseed oil, olive oil, evening primrose oil, flax seed oil,
palm oil, palm kernel oil, peanut oil, rapeseed oil, safflower oil,
sesame oil, sweet almond oil, rose hip oil, calendula oil,
chamomile oil, eucalyptus oil, juniper oil, safflower oil,
sandalwood oil, tea tree oil, sunflower oil, soybean oil, wheat
germ oil; animal oils such as shark liver oil, cod liver oil, whale
oil, beef tallow and butterfat; waxes such as beeswax, carnauba
palm wax, spermaceti and lanolin; fatty acids such as lauric acid,
myristic acid, palmitic, acid, stearic acid, oleic acid, behenic
acid; omega-3 fatty acids such as alpha-linolenic acid,
eicosapentaenoic acid, and docosahexaenoic acid; omega-6 fatty
acids such as linoleic acid and gamma-linolenic acid; aliphatic
alcohols such as lauryl, stearyl, cetyl, and oleyl alcohol; and
aliphatic esters such as isopropyl, isocetyl, or octadecyl
myristate, butyl stearate, hexyl laureate, diisopropyl ester of
adipic acid, or diisopropyl sebacate; and/or mixtures thereof.
Generally, the oils are refined and/or hydrogenated. Lipids include
monoglycerides, diglycerides, triglycerides, phospholipids, and
ceramides.
[0261] Suspending Agents
[0262] The compositions of the present invention may further
contain a suspending agent, preferably at concentrations effective
for suspending water-insoluble material in dispersed form in the
compositions or for modifying the viscosity of the composition.
Such concentrations can preferably range from about 0.1% to about
10%, more preferably from about 0.25% to about 5.0%. Suspending
agents useful herein include anionic polymers and nonionic
polymers. Useful herein are vinyl polymers such as cross linked
acrylic acid polymers with the CTFA name Carbomer, cellulose
derivatives and modified cellulose polymers such as methyl
cellulose, ethyl cellulose, nitro cellulose, sodium carboxymethyl
cellulose, crystalline cellulose, cellulose powder,
polyvinylpyrrolidone, polyvinyl alcohol, guar gum, hydroxypropyl
guar gum, arabia gum, galactan, carob gum, pectin, agar, starch
(rice, corn, potato, wheat), algae colloids (algae extract),
microbiological polymers such as dextran, succinoglucan, pulleran,
starch-based polymers such as carboxymethyl starch,
methylhydroxypropyl starch, alginic acid-based polymers such as
sodium alginate, alginic acid propylene glycol esters, acrylate
polymers such as sodium polyacrylate, polyethylacrylate,
polyacrylamide, polyethyleneimine, and inorganic water soluble
material such as bentonite, aluminum magnesium silicate, laponite,
hectonite, and anhydrous silicic acid. Actives aforementioned as
thickening agents can also be used herein as suspending agents.
[0263] Other optional suspending agents include crystalline
suspending agents which can be categorized as acyl derivatives,
long chain amine oxides, long chain acyl derivatives and mixtures
thereof. These preferred suspending agents include ethylene glycol
esters of fatty acids, alkanol amides of fatty acids, long chain
esters of long chain fatty acids (e.g., stearyl stearate, cetyl
palmitate, etc.); long chain esters of long chain alkanol amides
(e.g., stearamide diethanolamide distearate, stearamide
monoethanolamide stearate); and glyceryl esters (e.g., glyceryl
distearate, trihydroxystearin, tribehenin). Other suitable
suspending agents include primary amines having a fatty alkyl
moiety having at least about 16 carbon atoms, examples of which
include palmitamine or stearamine, and secondary amines having two
fatty alkyl moieties each having at least about 12 carbon atoms,
examples of which include dipalmitoylamine or di(hydrogenated
tallow)amine. Still other suitable suspending agents include
di(hydrogenated tallow)phthalic acid amide, and crosslinked maleic
anhydride-methyl vinyl ether copolymer.
[0264] Emulsifying Agents
[0265] Emulsifying agents include a wide variety of nonionic,
cationic, anionic, zwitterionic, and amphoteric surfactants such as
are known in the art and discussed below. The hydrophilic
surfactants (cationic, anionic, zwitterionic, amphoteric) useful
herein can contain a single surfactant, or any combination of
suitable surfactants. The exact surfactant (or surfactants) chosen
will depend upon the pH of the composition and the other components
present.
[0266] Useful nonionic surfactants include the condensation
products of alkylene oxides with fatty acids (i.e., alkylene oxide
esters of fatty acids), the condensation products of alkylene
oxides with 2 moles of fatty acids (i.e., alkylene oxide diesters
of fatty acids), the condensation products of alkylene oxides with
fatty alcohols (i.e., alkylene oxide ethers of fatty alcohols), the
condensation products of alkylene oxides with both fatty acids and
fatty alcohols [i.e., wherein the polyalkylene oxide portion is
esterified on one end with a fatty acid and etherified (i.e.,
connected via an ether linkage) on the other end with a fatty
alcohol]. Nonlimiting examples of these alkylene oxide derived
nonionic surfactants include ceteth-6, ceteth-10, ceteth-12,
ceteareth-6, ceteareth-10, ceteareth-12, steareth-6, steareth-10,
steareth-12, steareth-21, PEG-6 stearate, PEG-10 stearate, PEG-100
stearate, PEG-12 stearate, PEG-20 glyceryl stearate, PEG-80
glyceryl tallowate, PEG-10 glyceryl stearate, PEG-30 glyceryl
cocoate, PEG-80 glyceryl cocoate, PEG-200 glyceryl tallowate, PEG-8
dilaurate, PEG-10 distearate, and mixtures thereof. Still other
useful nonionic surfactants include polyhydroxy fatty acid amide
surfactants. An especially preferred surfactant corresponding to
the above structure is coconut alkyl N-methyl glucoside amide.
Preferred among the nonionic surfactants are those selected from
the group consisting of steareth-21, ceteareth-20, ceteareth-12,
sucrose cocotte, steareth-100, PEG-100 stearate, and mixtures
thereof. Other nonionic surfactants suitable for use herein include
sugar esters and polyesters, alkoxylated sugar esters and
polyesters, C1-C30 fatty acid esters of C1-C30 fatty alcohols,
alkoxylated derivatives of C1-C30 fatty acid esters of C1-C30 fatty
alcohols, alkoxylated ethers of C1-C30 fatty alcohols, polyglyceryl
esters of C1-C30 fatty acids, C1-C30 esters of polyols, C1-C30
ethers of polyols, alkyl phosphates, polyoxyalkylene fatty ether
phosphates, fatty acid amides, acyl lactylates, and mixtures
thereof. Nonlimiting examples of these emulsifiers include:
polyethylene glycol 20 sorbitan monolaurate (Polysorbate 20),
polyethylene glycol 5 soya sterol, Steareth-20, Ceteareth-20, PPG-2
methyl glucose ether distearate, Ceteth-10, Polysorbate 80, cetyl
phosphate, potassium cetyl phosphate, diethanolamine cetyl
phosphate, Polysorbate 60, glyceryl stearate, polyoxyethylene 20
sorbitan trioleate (Polysorbate 85), sorbitan monolaurate,
polyoxyethylene 4 lauryl ether sodium stearate, polyglyceryl-4
isostearate, hexyl laurate, PPG-2 methyl glucose ether distearate,
PEG-100 stearate, and mixtures thereof. Another group of non-ionic
surfactants useful herein are fatty acid ester blends based on a
mixture of sorbitan or sorbitol fatty acid ester and sucrose fatty
acid ester, the fatty acid in each instance being preferably
C8-C24, more preferably C10-C20. The preferred fatty acid ester
emulsifier is a blend of sorbitan or sorbitol C16-C20 fatty acid
ester with sucrose C10-C16 fatty acid ester, especially sorbitan
stearate and sucrose cocoate. This is commercially available from
ICI under the trade name Arlatone 2121.
[0267] Also useful herein are cationic surfactants, especially
dialkyl quaternary ammonium compounds. Nonlimiting examples of
these cationic emulsifiers include stearamidopropyl PG-dimonium
chloride phosphate, behenamidopropyl PG dimonium chloride,
stearamidopropyl ethyldimonium ethosulfate, stearamidopropyl
dimethyl (myristyl acetate) ammonium chloride, stearamidopropyl
dimethyl cetearyl ammonium tosylate, stearamidopropyl dimethyl
ammonium chloride, stearamidopropyl dimethyl ammonium lactate, and
mixtures thereof. Especially preferred is behenamidopropyl PG
dimonium chloride. Nonlimiting examples of quaternary ammonium salt
cationic surfactants include those selected from cetyl ammonium
chloride, cetyl ammonium bromide, lauryl ammonium chloride, lauryl
ammonium bromide, stearyl ammonium chloride, stearyl ammonium
bromide, cetyl dimethyl ammonium chloride, cetyl dimethyl ammonium
bromide, lauryl dimethyl ammonium chloride, lauryl dimethyl
ammonium bromide, stearyl dimethyl ammonium chloride, stearyl
dimethyl ammonium bromide, cetyl trimethyl ammonium chloride, cetyl
trimethyl ammonium bromide, lauryl trimethyl ammonium chloride,
lauryl trimethyl ammonium bromide, stearyl trimethyl ammonium
chloride, stearyl trimethyl ammonium bromide, lauryl dimethyl
ammonium chloride, stearyl dimethyl cetyl ditallow dimethyl
ammonium chloride, dicetyl ammonium chloride, dicetyl ammonium
bromide, dilauryl ammonium chloride, dilauryl ammonium bromide,
distearyl ammonium chloride, distearyl ammonium bromide, dicetyl
methyl ammonium chloride, dicetyl methyl ammonium bromide, dilauryl
methyl ammonium chloride, dilauryl methyl ammonium bromide,
distearyl methyl ammonium chloride, distearyl methyl ammonium
bromide, and mixtures thereof. Additional quaternary ammonium salts
include those wherein the C12 to C30 alkyl carbon chain is derived
from a tallow fatty acid or from a coconut fatty acid. The term
"tallow" refers to an alkyl group derived from tallow fatty acids
(usually hydrogenated tallow fatty acids), which generally have
mixtures of alkyl chains in the C16 to C18 range. The term
"coconut" refers to an alkyl group derived from a coconut fatty
acid, which generally have mixtures of alkyl chains in the C12 to
C14 range. Examples of quaternary ammonium salts derived from these
tallow and coconut sources include ditallow dimethyl ammonium
chloride, ditallow dimethyl ammonium methyl sulfate,
di(hydrogenated tallow) dimethyl ammonium chloride, di(hydrogenated
tallow) dimethyl ammonium acetate, ditallow dipropyl ammonium
phosphate, ditallow dimethyl ammonium nitrate,
di(coconutalkyl)dimethyl ammonium chloride,
di(coconutalkyl)dimethyl ammonium bromide, tallow ammonium
chloride, coconut ammonium chloride, and mixtures thereof. An
example of a quaternary ammonium compound having an alkyl group
with an ester linkage is ditallowyl oxyethyl dimethyl ammonium
chloride. More preferred cationic surfactants are those selected
from behenamidopropyl PG dimonium chloride, dilauryl dimethyl
ammonium chloride, distearyl dimethyl ammonium chloride, dimyristyl
dimethyl ammonium chloride, dipalmityl dimethyl ammonium chloride,
distearyl dimethyl ammonium chloride, stearamidopropyl PG-dimonium
chloride phosphate, stearamidopropyl ethyldiammonium ethosulfate,
stearamidopropyl dimethyl (myristyl acetate) ammonium chloride,
stearamidopropyl dimethyl cetearyl ammonium tosylate,
stearamidopropyl dimethyl ammonium chloride, stearamidopropyl
dimethyl ammonium lactate, and mixtures thereof. Still more
preferred cationic surfactants are those selected from
behenamidopropyl PG dimonium chloride, dilauryl dimethyl ammonium
chloride, distearyl dimethyl ammonium chloride, dimyristyl dimethyl
ammonium chloride, dipalmityl dimethyl ammonium chloride, and
mixtures thereof. A preferred combination of cationic surfactant
and structuring agent is behenamidopropyl PG dimonium chloride
and/or behenyl alcohol, wherein the ratio is preferably optimized
to maintain or to enhance physical and chemical stability,
especially when such a combination contains ionic and/or highly
polar solvents.
[0268] A wide variety of anionic surfactants can also be useful
herein. Nonlimiting examples of anionic surfactants include the
alkoyl isethionates, and the alkyl and alkyl ether sulfates. The
reaction products of fatty acids esterified with isethianonic acid
and neutralized, i.e., the alkoyl isethionates typically have the
formula RCOOCH2CH2SO3M wherein R is alkyl or alkenyl of from about
10 to about 30 carbon atoms, and M is a water-soluble cation such
as ammonium, sodium, potassium and triethanolamine. For example,
the fatty acids are derivated from coconut or palm kernel oil.
Nonlimiting examples of these isethionates include those alkoyl
isethionates selected from ammonium cocoyl isethionate, sodium
cocoyl isethionate, sodium lauroyl isethionate, sodium stearoyl
isethionate, and mixtures thereof. Also suitable are salts of fatty
acids, amids of methyl taurides. The alkyl and alkyl ether sulfates
typically have the respective formulae ROSO3M and RO(C2H40)xSO3M,
wherein R is alkyl or alkenyl of from about 10 to about 30 carbon
atoms, x is from about 1 to about 10, and M is a water-soluble
cation such as ammonium, alkanolamines such as triethanolamine,
monovalent metals, such as sodium and potassium, and polyvalent
metal cations such as magnesium and calcium. Preferably, R has from
about 8 to about 18 carbon atoms, more preferably from about 10 to
about 16 carbon atoms, even more preferably from about 12 to about
14 carbon atoms, in both the alkyl and alkyl ether sulfates. The
alkyl ether sulfates are typically made as condensation products of
ethylene oxide and monohydric alcohols having from about 8 to about
24 carbon atoms. The alcohols can be synthetic or they can be
derived from fats, e.g., coconut oil, palm kernel oil, tallow.
Lauryl alcohol and straight chain alcohols derived from coconut oil
or palm kernel oil are preferred. Such alcohols are reacted with
between about 0 and about 10, preferably from about 2 to about 5,
more preferably about 3, molar proportions of ethylene oxide, and
the resulting mixture of molecular species having, for example, an
average of 3 moles of ethylene oxide per mole of alcohol, is
sulfated and neutralized. Another suitable class of anionic
surfactants are the water-soluble salts of the organic, sulfuric
acid reaction products of the general formula R1-503-M, wherein R1
is chosen from the group including a straight or branched chain,
saturated aliphatic hydrocarbon radical having from about 8 to
about 24, preferably about 10 to about 16, carbon atoms; and M is a
cation described hereinbefore. Still other anionic synthetic
surfactants include the class designated as succinamates, olefin
sulfonates having about 12 to about 24 carbon atoms, and
beta-alkyloxy alkane sulfonates. Examples of these materials are
sodium lauryl sulfate and ammonium lauryl sulfate. Other anionic
surfactants suitable for use in the compositions are the
succinnates, examples of which include disodium
N-octadecylsulfosuccinnate; disodium lauryl sulfosuccinate;
diammonium lauryl sulfosuccinate; tetrasodium
N-0,2-dicarboxyethyfi-N-octadecylsulfosuccinnate; diamyl ester of
sodium sulfosuccinic acid; dihexyl ester of sodium sulfosuccinic
acid; and dioctyl esters of sodium sulfosuccinic acid. Other
suitable anionic surfactants include olefin sulfonates having about
10 to about 24 carbon atoms. In addition to the true alkene
sulfonates and a proportion of hydroxy-alkanesulfonates, the olefin
sulfonates can contain minor amounts of other materials, such as
alkene disulfonates depending upon the reaction conditions,
proportion of reactants, the nature of the starting olefins and
impurities in the olefin stock and side reactions during the
sulfonation process. Another class of anionic surfactants suitable
for use in the compositions is the beta-alkyloxy alkane sulfonate
class. Other anionic materials useful herein are soaps (i.e.,
alkali metal salts, e.g., sodium or potassium salts) of fatty
acids, typically having from about 8 to about 24 carbon atoms,
preferably from about 10 to about 20 carbon atoms. The fatty acids
used in making the soaps can be obtained from natural sources such
as, for instance, plant or animal-derived glycerides (e.g., palm
oil, coconut oil, soybean oil, castor oil, tallow, lard, etc.) The
fatty acids can also be synthetically prepared.
[0269] Amphoteric and zwitterionic surfactants are also useful
herein. Examples of amphoteric and zwitterionic surfactants which
can be used in the compositions of the present invention are those
which are broadly described as derivatives of aliphatic secondary
and tertiary amines in which the aliphatic radical can be straight
or branched chain and wherein one of the aliphatic substituents
contains from about 8 to about 22 carbon atoms (preferably C8-C18)
and one contains an anionic water solubilizing group, e.g.,
carboxy, sulfonate, sulfate, phosphate, or phosphonate. Examples
are alkyl imino acetates, and iminodialkanoates and aminoalkanoates
of the formulas RN[CH2)mCO2M]2 and RNH(CH2)mCO2M wherein m is from
1 to 4, R is a C8-C22 alkyl or alkenyl, and M is H, alkali metal,
alkaline earth metal ammonium, or alkanolammonium. Preferred
amphoteric surfactants for use in the present invention include
cocoamphoacetate, cocoamphodiacetate, lauroamphoacetate,
lauroamphodiacetate, and mixtures thereof. Also included are
imidazolinium and ammonium derivatives. Specific examples of
suitable amphoteric surfactants include sodium
3-dodecyl-aminopropionate, sodium 3-dodecylaminopropane sulfonate,
N-alkyltaurines such as the one prepared by reacting dodecylamine
with sodium isethionate; N-higher alkyl aspartic acids; and the
products sold under the trade name "Miranol". Other examples of
useful amphoterics include phosphates, such as coamidopropyl
PG-dimonium chloride phosphate (commercially available as Monaquat
PTC, from Mona Corp.). Zwitterionic surfactants suitable for use in
the composition are well known in the art, and include those
surfactants broadly described as derivatives of aliphatic
quaternary ammonium, phosphonium, and sulfonium compounds, in which
the aliphatic radicals can be straight or branched chain, and
wherein one of the aliphatic substituents contains from about 8 to
about 18 carbon atoms and one contains an anionic group such as
carboxy, sulfonate, sulfate, phosphate or phosphonate.
Zwitterionics such as betaines are preferred. Examples of betaines
include the higher alkyl betaines, such as coco dimethyl
carboxymethyl betaine, lauryl dimethyl carboxymethyl betaine,
lauryl dimethyl alphacarboxyethyl betaine, cetyl dimethyl
carboxymethyl betaine, cetyl dimethyl betaine (available as
Lonzaine 16SP from Lonza Corp.), lauryl bis-(2-hydroxyethyl)
carboxymethyl betaine, stearyl bis-(2-hydroxypropyl) carboxymethyl
betaine, oleyl dimethyl gamma-carboxypropyl betaine, lauryl
bis-(2-hydroxypropyl)alpha-carboxyethyl betaine, coco dimethyl
sulfopropyl betaine, stearyl dimethyl sulfopropyl betaine, lauryl
dimethyl sulfoethyl betaine, lauryl bis-(2-hydroxyethyl)
sulfopropyl betaine, and amidobetaines and amidosulfobetaines
(wherein the RCONH(CH2)3 radical is attached to the nitrogen atom
of the betaine), oleyl betaine (available as amphoteric Velvetex
OLB-50 from Henkel), and cocamidopropyl betaine (available as
Velvetex BK-35 and BA-35 from Henkel). Other useful amphoteric and
zwitterionic surfactants include the sultaines and hydroxysultaines
such as cocamidopropyl hydroxysultaine (available as Mirataine CBS
from Rhone-Poulenc), and the alkanoyl sarcosinates corresponding to
the formula RCON(CH3)CH2CH2CO2M wherein R is alkyl or alkenyl of
about 10 to about 20 carbon atoms, and M is a water-soluble cation
such as ammonium, sodium, potassium and trialkanolamine (e.g.,
triethanolamine), a preferred example of which is sodium lauroyl
sarcosinate.
[0270] Thickening Agents
[0271] Thickening agents suitable for inclusion in a composition
described herein include those agents commonly used as an excipient
or a carrier for topical application to increase the viscosity of
the formulation. Thickening agents may also be used to improve the
stability of the formulation and the product.
[0272] More specifically, such examples include but are not limited
to, acrylamides copolymer, agarose, amylopectin, bentonite, calcium
alginate, calcium carboxymethyl cellulose, carbomer, carboxymethyl
chitin, cellulose gum, dextrin, gelatin, hydrogenated tallow,
hydroxylethyl-cellulose, hydroxypropylcellulose, hydroxypropyl
starch, magnesium alginate, methylcellulose, microcrystalline
cellulose, pectin, various polyethylene glycol's, polyacrylic acid,
poly-methacrylic acid, polyvinyl alcohol, various polypropylene
glycols, sodium acrylates copolymer, sodium carrageenan, xanthan
gum, and/or yeast beta-glucan.
[0273] More generally, carboxylic acid polymers useful thickening
agents. Carboxylic acid polymers are cross-linked compounds
containing one or more monomers derived from acrylic acid,
substituted acrylic acids, and salts and esters of these acrylic
acids and the substituted acrylic acids, wherein the cross-linking
agent contains two or more carbon-carbon double bonds and is
derived from a polyhydric alcohol. Examples of commercially
available carboxylic acid polymers useful herein include the
carbomers, which are homopolymers of acrylic acid cross-linked with
allyl ethers of sucrose or pentaerytritol. The carbomers are
available as the Carbopol.RTM. 900 series from B.F. Goodrich (e.g.,
Carbopol.RTM. 954). In addition, other suitable carboxylic acid
polymeric agents include copolymers of C10-30 alkyl acrylates with
one or more monomers of acrylic acid, methacrylic acid, or one of
their short chain (i.e., C1-4 alcohol) esters, wherein the
cross-linking agent is an allyl ether of sucrose or pentaerytritol.
These copolymers are known as acrylates/C10-30 alkyl acrylate
crosspolymers and are commercially available as Carbopol.RTM. 1342,
Carbopol.RTM. 1382, Pemulen TR-1, and Pemulen TR-2, from B.F.
Goodrich. Examples of preferred carboxylic acid polymer thickeners
useful herein include those selected from carbomers,
acrylates/C10-30 alkyl acrylate crosspolymers, and mixtures
thereof.
[0274] Moreover, a wide variety of polysaccharides are useful
herein as thickening agents. Non-limiting examples of
polysaccharide gelling agents include those selected from
cellulose, carboxymethyl hydroxyethylcellulose, cellulose acetate
propionate carboxylate, hydroxyethylcellulose, hydroxyethyl
ethylcellulose, hydroxypropylcellulose, hydroxypropyl
methylcellulose, methyl hydroxyethylcellulose, microcrystalline
cellulose, sodium cellulose sulfate, and mixtures thereof. Also
useful herein are the alkyl substituted celluloses. In these
polymers, the hydroxy groups of the cellulose polymer is
hydroxyalkylated (preferably hydroxyethylated or hydroxypropylated)
to form a hydroxyalkylated cellulose which is then further modified
with a C10-30 straight chain or branched chain alkyl group through
an ether linkage Typically these polymers are ethers of C10-30
straight or branched chain alcohols with hydroxyalkylcelluloses.
Examples of alkyl groups useful herein include those selected from
stearyl, isostearyl, lauryl, myristyl, cetyl, isocetyl, cocoyl
(e.g., alkyl groups derived from the alcohols of coconut oil),
palmityl, oleyl, linoleyl, linolenyl, ricinoleyl, behenyl, and
mixtures thereof. Preferred among the alkyl hydroxyalkyl cellulose
ethers is the material given the CTFA designation cetyl
hydroxyethylcellulose, which is the ether of cetyl alcohol and
hydroxyethylcellulose. This material is sold under the trade name
Natrosol.RTM. CS Plus from Aqualon Corporation (Wilmington, Del.).
Additional examples can be found in The International Cosmetic
Ingredient Dictionary and Handbook, the Cosmetic Bench
Reference--Directory of Cosmetic Ingredients, the books provided by
the United States Pharmacopeia (USP) and the National Formulary
(NF), and other references for cosmetic and pharmaceutical
ingredients known in the art. Other useful polysaccharides include
scleroglucans which are a linear chain of (1-3) linked glucose
units with a (1-6) linked glucose every three units, a commercially
available example of which is Clearogel.TM. CS11 from Michel
Mercier Products Inc. (Mountainside, N.J.).
[0275] Other thickening and gelling agents useful herein include
materials which are primarily derived from natural sources.
Non-limiting examples of these gelling agent gums include acacia,
agar, algin, alginic acid, ammonium alginate, amylopectin, calcium
alginate, calcium carrageenan, carnitine, carrageenan, dextrin,
gelatin, gellan gum, guar gum, guar hydroxypropyltrimonium
chloride, hectorite, hyaluronic acid, hydrated silica,
hydroxypropyl chitosan, hydroxypropyl guar, karaya gum, kelp,
locust bean gum, natto gum, potassium alginate, potassium
carrageenan, propylene glycol alginate, sclerotium gum, sodium
carboyxmethyl dextran, dextran sulfate, sodium carrageenan,
tragacanth gum, xanthan gum, and/or mixtures thereof. In addition,
the compositions of the present invention can also optionally
contain polyacrylamide polymers, especially nonionic polyacrylamide
polymers including substituted branched or unbranched polymers.
More preferred among these polyacrylamide polymers is the nonionic
polymer given the CTFA designation polyacrylamide and isoparaffin
and laureth-7, available under the trade name Sepigel 305 from
Seppic Corporation (Fairfield, N.J.). Other polyacrylamide polymers
useful herein include multi-block copolymers of acrylamides and
substituted acrylamides with acrylic acids and substituted acrylic
acids.
[0276] Preferred compositions of the present invention include a
thickening agent selected from carboxylic acid polymers,
cross-linked polyacrylate polymers, polyacrylamide polymers, and
mixtures thereof, more preferably selected from carboxylic acid
polymers, polyacrylamide polymers, and mixtures thereof.
[0277] Penetration Enhancers
[0278] Penetration enhancers are the substances that facilitate the
absorption of penetrant through the skin or mucosal membranes by
temporarily diminishing the impermeability of the skin or,
respectively, the mucosa. Ideally, these materials should be
pharmacologically inert, nontoxic, nonirritating, non-allergenic,
compatible with Octanoyl Carnosine, odorless, tasteless, colorless,
and inexpensive and have good solvent properties. The enhancer
should not lead to the significant loss of body fluids,
electrolytes, and other endogenous materials, and skin or mucosa
should regain its barrier properties on its removal within an
acceptable period of time. No single penetration enhancer can
possess all the required properties. However, many enhancers
exhibit many of these attributes, and they have been described (for
example as reviewed in Drug Development and Industrial Pharmacy
2000, 26, 1131-1140) or are being currently researched.
[0279] Anti-Histamines
[0280] Anti-histamines, also called histamine antagonists, are
substances that inhibit the action of histamine by blocking it from
attaching to histamine receptors; or by inhibiting the enzymatic
activity of histidine decarboxylase, catalyzing the transformation
of histidine into histamine; or similar Examples of anti-histamines
are acrivastine, azelastine, brompheniramine, buclizine,
bromodiphenhydramine, carbinoxamine, cetirizine, chlorpromazine,
cyclizine, chlorpheniramine, chlorodiphenhydramine, cimetidine,
clemastine, cyproheptadine, desloratadine, dexbrom-pheniramine,
deschlorpheniramine, dexchlorpheniramine, dimenhydrinate,
dimetindene, diphenhydramine, doxylamine, ebastine, embramine,
famotidine, fexofenadine, lafutidine, levocetirizine, loratadine,
meclozine, mirtazapine, nizatidine, olopatadine, orphenadrine,
phenindamine, pheniramine, phenyltoloxamine, promethazine,
pyrilamine, quetiapine, ranitidine, roxatidine, rupatadine,
tripelennamine, and triprolidine.
[0281] The choice of additional substances to be included in the
composition is made depending on the constraints relating to
Octanoyl Carnosine (or one or more derivatives thereof) (e.g.,
stability, solubilization, etc.), if enhanced and/or additional
benefits and properties (e.g., anti-acne, anti-microbial,
anti-wrinkle, skin lightening, anti-redness, antioxidant, skin
protectant, sunscreen, hair growth, anti-inflammatory, emolliant,
moisturization, enhanced skin penetration, etc.) of the composition
are desired, and, where applicable, the use subsequently envisaged
for the composition.
[0282] As mentioned, the compositions of the invention may include
one or more additional substances, various, conventional or not,
which will provide some benefit to the object of the composition.
More specifically, the combination of Octanoyl Carnosine (or one or
more derivatives thereof) with selected additional substances may
lead to an enhanced efficacy as compared to the use of Octanoyl
Carnosine (or one or more derivatives thereof) alone. The enhanced
efficacy can be additive (the sum of efficacies of the individual
agents alone), or it can be synergistic (larger than the sum of
efficacies of the individual agents alone). Of course, a decision
to include an additional ingredient or substance and the choice of
a specific ingredient or substance depends on the specific use of
the composition and the product formulation.
[0283] For example, the use of one or more suitable anti-wrinkling
substance (e.g., retinoic acid, retinol, transforming growth factor
beta-1, selected peptides, etc.) will increase the clinical
efficacy (e.g., reduced skin wrinkles) of the composition
containing Octanoyl Carnosine (or one or more derivatives thereof)
after topical administration; the use of one or more suitable
emollient substance (e.g., octyldodecanol, etc.) will increase the
clinical efficacy (e.g., improved skin feel or sensations) of the
composition containing Octanoyl Carnosine (or one or more
derivatives thereof) after topical administration; the use of one
or more suitable humectant substance (e.g., glycerin, hyaluronic
acid, etc.) will increase the clinical efficacy (e.g., increased
skin moisturization) of the composition containing Octanoyl
Carnosine (or one or more derivatives thereof) after topical
administration; the use of one or more suitable skin penetration
enhancer substance (e.g., propylene glycol, butylene glycol,
ethanol, oleic acid, lauric acid, palmitic acid, isopropyl
palmitate, DMSO, sodium lauryl sulfate, Azone.RTM., etc.) for
Octanoyl Carnosine (or one or more derivatives thereof) will
increase the clinical efficacy (e.g., reduced skin wrinkles) of the
composition containing Octanoyl Carnosine (or one or more
derivatives thereof) after topical administration; the use of one
or more suitable anti-inflammatory substance (e.g., bisabolol,
glycyrrhetinic acid, linoleic acid, borage seed oil, wheat germ
oil, etc.) will increase the clinical efficacy (e.g., reduced
irritation or redness of skin or mucosa) of the composition
containing Octanoyl Carnosine (or one or more derivatives thereof)
after topical administration; the use of one or more suitable
topical anesthetic substance (e.g., lidocaine, pramoxine
hydrochloride, etc.) will increase the clinical efficacy (e.g.,
reduced local pain) of the composition containing Octanoyl
Carnosine (or one or more derivatives thereof) after topical
administration; and the use of one or more suitable topical
anti-histamine substance (e.g., diphenhydramine, etc.) will
increase the clinical efficacy (e.g., reduced local itch) of the
composition containing Octanoyl Carnosine (or one or more
derivatives thereof) after topical administration.
Carriers and Excipients
[0284] The compositions of the present invention can also contain
one or more carriers and/or excipients acceptable for a mode of
administration (i.e., for topical application and/or for
subcutaneous administration). Those skilled in the art will be able
to routinely select an appropriate carrier and/or excipient for the
mode of administration. Depending in the use and the way of
administration, the compositions of the present invention can also
contain a carrier and/or excipient acceptable for injection,
implantation, or subcutaneous placement.
[0285] The carrier and/or excipient can be in a wide variety of
forms. Non-limiting examples of suitable carriers and/or excipients
include simple solutions (water or oil based), emulsions,
dispersions, multi-phase systems, semi-solid forms, solid forms
(powder, sticks, patches), skin masks, tissues, foams, and
aerosols. For example, emulsion carriers and/or excipients can
include, but are not limited to, oil-in-water, silicone-in-water,
water-in-oil, water-in-silicone, water-in-oil-in-water,
oil-in-water-in-oil, and oil-in-water-in-silicone emulsions.
[0286] Depending upon the desired product form, preferred carriers
and/or excipients can contain an emulsion such as oil-in-water
emulsions (e.g., silicone-in-water) and water-in-oil emulsions,
(e.g., water-in-silicone emulsions). In one example, oil-in-water
emulsions are especially preferred. Emulsions according to the
present invention can contain an aqueous phase and a lipid or oil.
Lipids and oils may be derived from animals, plants, or petroleum
and may be natural or synthetic (e.g., man-made).
[0287] Preferred emulsions can also contain a humectant, such as
glycerin. Emulsions can further contain from about 0.1% to about
25%, more preferably from about 0.2% to about 10%, of an
emulsifying agent (emulsifier), based on the weight of the
composition. Emulsifier agents may be nonionic, anionic, or
cationic. Suitable emulsifiers are disclosed for example in
McCutcheon's Detergents and Emulsifiers, North American Edition,
pages 317-324 (1986).
[0288] Suitable emulsions may have a wide range of viscosities,
depending on the desired product form. The compositions of the
present invention can be in the form of pourable liquids,
semi-solids, to highly viscous systems (e.g., solids) under ambient
conditions.
[0289] Any of the compositions can contain an aqueous carrier
and/or excipient, which is typically present at a level of from
about 20% to about 99%, preferably from about 60% to about 90%. The
aqueous carrier and/or excipient may contain water, or a miscible
mixture of water and organic solvent (e.g., alcohols, including but
not limited to ethanol, glycerin, propylene glycol, butylene
glycol, other glycols, etc.), but preferably contain water with
significant lower or no concentrations of organic solvent, except
as otherwise incidentally incorporated into the composition as
minor ingredients of other essential or optional components.
[0290] The compositions of the subject invention, including but not
limited to solutions, lotions, serums and creams, may contain an
acceptable emollient. Such compositions preferably contain from
about 1% to about 80% of the emollient. As used herein, "emollient"
refers to a material useful for the prevention or relief of
dryness, as well as for the protection of the skin. A wide variety
of suitable emollients is known and may be used herein. In addition
to the examples of emollients provided above, Sagarin, Cosmetics,
Science and Technology, 2nd Edition, Vol. 1, pp. 32-43 (1972)
contains numerous other examples of materials suitable as an
emollient. A preferred emollient is glycerin. Glycerin is
preferably used in an amount of from or about 0.001% to or about
80%, more preferably from or about 0.01% to or about 25%, still
more preferably from or about 0.1% to or about 10%, and even more
preferably from or about 2% to or about 5%.
[0291] Lotions, serums and creams according to the present
invention generally contain a carrier and/or excipient and one or
more emollients. Lotions and creams typically contain from about 1%
to about 50%, preferably from about 1% to about 20%, of emollient;
from about 50% to about 90%, preferably from about 60% to about
80%, water; and, optionally, additional substances in amounts
sufficient to provide additional benefits. Creams are generally
thicker than lotions and serums due to higher levels of emollients,
higher levels of thickeners, and/or differences in the emulsifying
system.
[0292] Ointments of the present invention may contain a simple base
of animal or vegetable oils or semi-solid hydrocarbons such as
petrolatum; absorption ointment bases which absorb water to form
emulsions; or water soluble carriers, e.g., a water soluble
solution carrier. Ointments may further contain a thickening agent,
such as described above and in Sagarin, Cosmetics, Science and
Technology, 2nd Edition, Vol. 1, pp. 72-73 (1972), and/or an
emollient. For example, an ointment may contain from about 2% to
about 10% of an emollient; from about 0.1% to about 2% of a
thickening agent as well as one or more additional substances(s) in
amounts sufficient to provide additional benefits.
[0293] Compositions of this invention useful for cleansing
("cleansers") can be formulated with a suitable carrier, e.g., as
described above, and preferably contain from about 1% to about 30%,
more preferably from about 5% to about 10%, of an acceptable
surfactant. The surfactant is suitably selected from anionic,
nonionic, zwitterionic, amphoteric and ampholytic surfactants, as
well as mixtures of these surfactants. Such surfactants are well
known to those skilled in the detergency art. Examples of a broad
variety of surfactants useful herein are described above and in
McCutcheon's Detergents and Emulsifiers, North American Edition
(1986), published by Allured Publishing Corporation. The cleansing
compositions can optionally contain, at their art-established
levels, other materials which are conventionally used in cleansing
compositions.
[0294] As used herein, the term "foundation" refers to a liquid,
semi-liquid, semi-solid, or solid skin cosmetic which includes, but
is not limited to lotions, creams, gels, pastes, cakes, and the
like. Typically the foundation is used over a large area of the
skin, such as over the face, to provide a particular look.
Foundations are typically used to provide an adherent base for
color cosmetics such as rouge, blusher, powder and the like, and
tend to hide skin imperfections and impart a smooth, even
appearance to the skin. Foundations of the present invention
include a dermatologically acceptable carrier and may include
conventional ingredients such as oils, colorants, pigments,
emollients, fragrances, waxes, stabilizers, and the like.
[0295] The compositions of the present invention can also contain a
liquid that is acceptable for injection to and/or under the skin if
the composition is to be injected. Any suitable acceptable liquid
as known in the art or otherwise can be used.
Composition Preparation
[0296] The compositions of the present invention are generally
prepared by conventional methods such as are known in the art of
making compositions suitable for topical application. Such methods
can typically be conducted in one or more steps, with or without
heating, cooling, and the like.
[0297] In addition, the compositions of the present invention can
also be prepared by conventional methods such as are known in the
art of making compositions suitable for injections.
[0298] As used herein, a "formulation" is a mixture prepared
according to a specific procedure.
[0299] The physical form of the compositions according to the
invention is not important. They may be in any galenic form such
aerosols, creams, lotions, milk or cream ointments, gels,
emulsions, dispersions, solutions, suspensions, cleansers,
foundations, anhydrous preparations (sticks, in particular lip
balm, body and bath oils), shower and bath gels, shampoos and scalp
treatment lotions, cream or lotion for care of skin or hair, gel or
solution for care of skin or hair, cream or lotion for care of the
genitals (e.g., vulva, vagina, penis, scrotum), gel or solution for
care of genitals, make-up removing lotions or creams, sunscreen
lotions, milks, artificial suntan lotions; pre-shave, shave or
after shave creams, foams, gels or lotions; make-up, lipsticks,
mascaras or nail varnishes; skin essences, serums; adhesive or
absorbent materials, skin masks; tissues; hydrating patches,
transdermal patches, iontophoretic patches, microneedle patches;
powders; emollient lotion, sprays, oils for the body and the bath,
foundation tint bases, pomade, colloid, compact or solid
suspension, pencil, sprayable or brossable formulation, blush,
rouge, eyeliner, lip liner, lip gloss, facial or body powder,
mousse or styling gels, nail conditioner, lip balms, skin
conditioners, anorectal creams, hygiene cream, moisturizers, hair
sprays, hair conditioners, soaps, body exfoliants, astringents,
depilatories and permanent waving solutions, anti-dandruff
formulations, anti-hair loss formulations, anti-sweat and
anti-perspirant formulations, nose sprays; and so on.
[0300] These compositions can also be presented in the form of
lipsticks intended to apply color or to protect the lips from
cracking, or of make-up products for the eyes or tints and tint
bases for the face. Compositions in accordance with the invention
include cosmetics, personal care products, feminine products, male
products, hygiene products, and dermatological or pharmaceutical
preparations.
[0301] The compositions of the present invention may also be
applied on animal skin when wounds or defects or disorders of
animal skin affecting the extracellular matrix are present.
[0302] The compositions according to the present invention may be
prepared in the form of solution, dispersion, emulsion, paste, or
powder, individually or as a premix or in vehicles individually or
as a premix in vectors such as macro-, micro-, or nanocapsules,
macro-, micro- or, nanospheres, liposomes, oleosomes, cubosomes;
macro-, micro-, or nanoparticles; or macro-, micro or nanosponges;
or macro-, micro-, and nanocapsules; or macro-, micro- or
nanospheres; micro- or nano-emulsions; or adsorbed onto tip of
needles; or adsorbed onto microneedles or onto microneedle arrays;
or adsorbed to organic polymer powders, talcs, bentonites, or other
inorganic or organic supports.
[0303] Furthermore, the compositions according to the present
invention may be used in any form whatsoever, in a form bound to or
incorporated in or absorbed in or adsorbed on macro-, micro-, and
nanoparticles; or macro-, micro or nanosponges; or macro-, micro-,
and nanocapsules; or macro-, micro- or nanospheres; or adsorbed
(e.g., by coating) onto microneedle patches or arrays (such as
described by Amen M. et al., Pharm Res 2010, 27: 303-313); for the
treatment of textiles, natural or synthetic fibers, wools, and any
materials that may be used for clothing or underwear for day or
night intended to come into contact with the skin, handkerchiefs or
cloths, to exert their effect via this skin/textile contact and to
permit continuous topical deliver.
[0304] The compositions according to the present invention may also
be prepared or used in a form of a device (e.g., medical device,
combination between drug and medical device). Preferred devices
include, but are not limited to, devices for overcoming biological
barriers such as ultrasound devices (i.e., sonophoresis,
sonoporation, acoustic ablation), electric devices (iontophoresis,
electroporation), high pressure devices (i.e., liquid injection,
powder injection), microneedles (i.e., solid, hollow, degradable,
coated), thermal and optical devices (i.e., light, infrared, laser,
radio-frequency), other physical devices reducing the skin barrier
(i.e., plasma devices, micro-dermabrasion, dermabrasion, suction
devices, macro-needle devices, etc.), devices reducing the skin
barrier by chemical means (i.e., chemical exfoliating devices, skin
corrosion (e.g., using NaOH) devices), and/or any combination or
combination device thereof. Some example of methods and devices for
overcoming biological barriers have been described in Advanced Drug
Delivery Reviews 2013, 65, 100-103 (incorporated herein as
reference).
[0305] In addition, the compositions according to the present
invention may be used in any form intended to be placed into the
skin or mucosal tissue, or under the skin or mucosal tissue (e.g.,
by injection, implantation, or subcutaneous placement).
Method of Treatment
[0306] The present invention concerns compositions for their
application as a cosmetic, personal care, or a medicinal
product.
[0307] The composition according to the invention can be applied
topically onto any areas of the face, neck, neckline, decollete,
scalp, hand, palm, arm, leg, foot, sole, chest, breast, back,
abdomen, buttock, vulva, or penis and scrotum, anus, and/or any
other skin areas of the human body.
[0308] Further, the composition according to the invention can be
also applied locally or topically onto any areas of the eye, mouth,
nose; breast nipples, vulva, vagina and introitus; or penis and
scrotum; rectum, and/or any other mucosal areas of the human
body.
[0309] Furthermore, the composition according to the invention can
also be applied locally or topically to other surfaces of the human
body, including hair and nail, or any wound, scar, or skin and
mucosal surface areas affected by atrophy, or other conditions,
disorders and diseases associated with changes in extracellular
matrix components.
[0310] In addition, the compositions according to the present
invention may also be applied by injection, implantation, or
subcutaneous placement.
[0311] For example, the compositions described herein can be
applied using a syringe, a micro-cannula, a patch, an iontophoretic
patch, microneedles, and/or a microneedle array or patch. In
addition, the composition can be also applied in conjunction (i.e.,
before, after, or simultaneously) with the use of other skin
devices changing the penetration characteristics of skin such as,
for example, laser, light, infrared, radiofrequency, ultrasound,
electroporation, sonophoresis, thermal, plasma, and/or high
pressure devices, and/or any combination(s) (including combination
devices) thereof. Any other commonly used means of administration
can also be utilized.
[0312] In addition, the compositions according to the present
invention may also be applied in animals.
[0313] In one example, the present invention concerns cosmetic
treatment methods to improve the general state of the skin
involving topical application of an effective amount of the
composition as defined above to the skin. More specifically, these
methods can be used to prevent and/or treat the signs of intrinsic
and extrinsic skin aging; to prevent and/or treat skin slackening
and/or improve tone and/or firmness and/or elasticity of the skin;
to prevent and/or treat skin atrophy and/or improve the density of
the dermis and epidermis; to give or return volume to the dermis
and epidermis; to prevent and/or treat skin dehydration; to prevent
and/or treat skin roughness; to prevent and/or treat cellulite, to
prevent and/or treat stretch marks, to reduce expansion and/or
prevent the development of adipose tissue within the hypodermis; to
lighten and/or whiten the skin; to prevent and/or treat glycation
of molecules in the skin; and/or to prevent and/or treat
degradation of the skin due to the effects of oxidation.
[0314] The present invention also provides methods to improve the
general state of the atrophic tissue involving topical application
of an effective amount of the composition as defined above to the
tissue. More specifically, such methods can be used to restore
damaged skin; to restore skin after cosmetic and dermatological
procedures; to prevent and/or treat atrophy of the female genitals,
to prevent and/or treat vulvovaginal atrophy; to prevent and/or
treat skin conditions and disorders related to menopause; to
prevent and/or treat skin conditions and disorders associated with
reduced estrogen levels in females; to prevent and/or treat
vulvodynia; to prevent and/or treat vulvar lichen sclerosus; to
prevent and/or treat vulvar dermatoses; to prevent and/or treat the
signs of intrinsic and extrinsic aging of the female genitals;
and/or to prevent and/or treat the signs of intrinsic and extrinsic
aging of male genitals.
[0315] Some benefits (e.g., moisturization, soothing, calming,
tightening, smooth feel, etc.) can be noticed within a few hours to
a few days after topically applying the compositions according to
the present invention on the affected human skin or human tissue
(for example vulva and/or vagina). However, it takes generally at
least 30 days to notice benefits (e.g., anti-aging, wrinkle
reducing, skin lightening, anti-redness, improving atrophy, wound
healing, etc.). Thereby, the composition should be applied to the
affected human skin or human tissue at least once to twice a
day.
[0316] Determination of an effective dose (e.g., therapeutically,
cosmetically, pharmaceutically, and/or medicinally effective dose)
of any of the compositions of the instant invention is within the
routine level of skill in the art.
Kits and Dosage Forms
[0317] According to the invention, products or devices with several
compartments or kits (having one or more containers) may be
proposed to apply the compositions of the invention. By way of
non-limiting example, a first compartment or container having a
composition including the Octanoyl Carnosine, the selected
tri-peptides, and/or tetra-peptides (or one or more derivatives
thereof), and one or more additional substances (e.g., one or more
biologically active ingredients and/or one or more inactive
ingredients such as an excipient and/or a carrier) in a second
compartment or container, the compositions contained in the said
first and second compartments in this case being considered to be a
combination composition for simultaneous, separate or step-wise use
in time, particularly in any one of the treatments defined above.
Alternatively, kits according to the invention may include the
components of the compositions in separate compartments or
containers or certain components can be in the same compartments or
containers while others are in separate compartments or containers.
Such kits will also preferably include instructions for use.
[0318] Any of the compositions described herein may be supplied in
dosage unit form for ease of administration and uniformity of
dosage. Dosage unit form as used herein refers to physically
discrete units suited as unitary dosages for the mammalian subjects
to be treated; each unit containing a predetermined quantity of
Octanoyl Carnosine (and one or more additional active ingredients)
calculated to produce the desired cosmetic, personal care or
therapeutic effect in association with the required cosmetic and/or
pharmaceutical carrier(s). The specification for the dosage unit
forms of the invention are dictated by and directly dependent on
(a) the unique characteristics of the compositions and the
particular maintenance, therapeutic or prophylactic effect to be
achieved, and (b) the limitations inherent in the art of
compounding Octanoyl Carnosine (and one or more additional active
ingredients) for the treatment of individuals.
[0319] The unit dosage form is any of a variety of forms,
including, for example, but not limited to, a solution, any
semi-solid form, a capsule, a bag, a tablet, a single pump on an
aerosol or a vial. The quantity of active ingredient(s) in a unit
dose of composition is an effective amount and is varied according
to the particular treatment involved.
[0320] One skilled in the art will appreciate that it is sometimes
necessary to make routine variations to the dosage depending on the
age and condition of the patient. The dosage will also depend on
the route of administration.
EXAMPLES
[0321] Examples of methods of chemical synthesis of Octanoyl
Carnosine, Palmitoyl-GHK, and GEKG, as well as suitable
compositions and their preparation according to the present
invention will be described hereafter. These compositions and their
preparation are representative, but do not restrict, the scope of
the invention.
[0322] The Examples set forth herein are meant to exemplify the
various aspects of carrying out the invention and are not intended
to limit the invention in any way. Unless otherwise specified, it
is to be understood that the concentrations of the ingredients in
the compositions of the invention are in weight percentages (w %),
based on the total weight of the composition. All measurements are
performed at 25.degree. Celsius unless stated otherwise.
[0323] The following examples describe and demonstrate various
aspects within the scope of the present invention. The examples are
only given for illustrative purposes and should not be considered
to be restrictive to this invention. Additionally for illustrative
purposes several compositions and their use for topical
administration will be described.
Example 1
Example of Synthesis of Peptide Octanoyl Carnosine
[0324] The di-peptide Octanoyl Carnosine can be prepared according
to standard procedures in peptide chemistry using solid-phase
synthesis or liquid-phase synthesis. As an example, one way of the
synthesis and purification of Octanoyl Carnosine
(Octanoyl-beta-Ala-His-OH) in its acetate salt form by solid-phase
synthesis is illustrated in FIG. 1.
[0325] The Octanoyl Carnosine synthesized can be purified using
standard methods (e.g., crystallization (as shown in this Example),
thin layer chromatography, column chromatography, preparative HPLC,
liquid-liquid extraction, etc.). Its identity can be confirmed by
ESI-MS, and its purity can be determined by HPLC. Other analytical
methods for peptides or lipoamino-peptides known in the art can
also be used to determine the identity and purity.
Example 2
Example of Synthesis of Palmitoyl-GHK
[0326] The tri-peptide Palmitoyl-GHK can be prepared according to
standard procedures in peptide chemistry using solid-phase
synthesis or liquid-phase synthesis. As an example, one way of the
synthesis and purification of Palmitoyl-GHK
(Palmitoyl-Gly-His-Lys-OH) in its acetate salt form by solid-phase
synthesis is illustrated in FIG. 2.
[0327] The Palmitoyl-GHK synthesized can be purified using standard
methods (e.g., crystallization (as realized in this Example), thin
layer chromatography, column chromatography, preparative HPLC,
liquid-liquid extraction, etc.). Its identity can be confirmed by
ESI-MS, and its purity can be determined by HPLC. Other analytical
methods for peptides or lipoamino-peptides known in the art can
also be used to determine the identity and purity.
Example 3
Example of Synthesis of Peptide GEKG
[0328] Peptide GEKG (NH.sub.2-Gly-Glu-Lys-Gly-OH) (SEQ ID NO:1) can
be prepared according to standard procedures in peptide chemistry
using solid-phase synthesis or liquid-phase synthesis known in the
art. (See, e.g., Merrifield, J. AM. CHEM. SOC. 85:2149-54 (1963)
(incorporated herein by reference)).
[0329] The GEKG synthesized can be purified using standard methods
(e.g., crystallization (as realized in this Example), thin layer
chromatography, column chromatography, preparative HPLC,
liquid-liquid extraction, etc.). Its identity can be confirmed by
ESI-MS, and its purity can be determined by HPLC. Other analytical
methods for peptides or lipoamino-peptides known in the art can
also be used to determine the identity and purity.
Example 4
Preparation of Compositions
[0330] This Example illustrates the preparation of a series of
compositions suitable for topical application in accordance to the
present invention. These compositions may contain Octanoyl
Carnosine (or one or more derivatives thereof) alone in a carrier
acceptable for topical applications; the combination of both
Octanoyl Carnosine and GEKG (or one or more derivatives thereof);
the combination of both Octanoyl Carnosine and Palmitoyl-GHK (or
one or more derivatives thereof); and/or the combination of
Octanoyl Carnosine, GEKG and Palmitoyl-GHK (or one or more
derivatives thereof). In addition, any of the compositions may also
include one or more additional substances and/or one or more
acceptable carriers and/or excipients suitable for topical
applications.
[0331] The peptides Octanoyl Carnosine, Palmitoyl-GHK, GEKG (as
well as their derivatives) can be synthesized in accordance with
the methods described in Examples 1, 2 and 3, respectively.
Alternatively, they can be obtained by different processes of
synthesis and purifications known in the art. Octanoyl Carnosine,
Palmitoyl-GHK, GEKG (or their derivatives) may be also obtained
from any peptide manufacturer or supplier; either in solid form
(i.e., as salt at high purity; e.g., larger than 75%, ideally
larger than 90%, and more ideally larger than 95%), or already
solublized in a simple and acceptable solvent, excipient, and/or
carrier suitable for topical application (e.g., water, glycerin,
propylene glycol, butylene glycol, hexylene glycol, low molecular
weight PEGs such as PEG-400, and/or any mixtures thereof; with or
without suitable surfactants helping to solubilize and stabilize
the peptides; and/or with or without anti-microbial preservatives;
and/or with or without chemical stabilizers including antioxidants
or iron chelators).
[0332] Preferably, Octanoyl Carnosine (or one or more derivatives
thereof) is present in the compositions in proportions of between
0.0001% and 10% of the total weight of the composition, more
preferably between 0.001% and 5%, and even more preferably between
0.005% and 1%, depending on the use of the composition and the more
or less potent effect sought. Thus, one of the preferred
composition contains Octanoyl Carnosine (or one or more derivatives
thereof) in proportions of between 0.005% and 1% in a carrier
acceptable for topical application.
[0333] The compositions according to the invention can be prepared
by adding Octanoyl Carnosine (or one or more derivatives thereof),
either in its solid form, or solubilized in a suitable solvent,
excipient, or carrier suitable for topical application, at any
stage of the preparation process of the composition. Typically,
however, Octanoyl Carnosine (or one or more derivatives thereof) is
added as part of the aqueous phase during the preparation process
of the composition. For instance in case of oil-in-water type
compositions, hydrogels, and aqueous systems, Octanoyl Carnosine
(or one or more derivatives thereof) can be added as the last step
to the previously prepared bulk of the composition by mixing or
homogenizing (e.g., using a mixer or homogenizer) Octanoyl
Carnosine (or one or more derivatives thereof), which has been
previously solubilized or incorporated into a solvent, excipient,
and/or carrier system, to the bulk of the composition at lower
temperatures (e.g., typically below 60.degree. C.; and ideally
between about 20.degree. to 40.degree. C.).
[0334] For example, a solvent, excipient, and/or carrier system
suitable for topical application and able to solubilize or
incorporate Octanoyl Carnosine (or one or more derivatives thereof)
can contain one or more of the following ingredients: water,
glycerin, propylene glycol, propanediol, butylene glycol, hexylene
glycol, low molecular weight PEGs such as PEG-400, cyclodextrins,
ethanol, and/or any mixtures thereof. Additionally, this solvent or
carrier system may further contain suitable surfactants (e.g.,
Tween 20, Tween 80, etc.) to help solubilize and stabilize the
peptides, and/or anti-microbial preservatives to help prevent
microbial growth and contamination, and/or antioxidants or iron
chelators (e.g., EDTA, etc.) as chemical stabilizers. Water,
glycerin, propylene glycol, propanediol, butylene glycol,
cyclodextrins, and/or any mixtures thereof, are the preferred
ingredients.
[0335] In most instances, the additional ingredients will include a
cosmetic, dermatologically or pharmaceutically acceptable carrier
either alone or in combination with other additional ingredients.
The amounts of additional ingredients may range from about 90% to
about 99.9999%, preferably from about 95% to about 99.999%, more
preferably from about 99% to about 99.999%, of the composition. In
short, the percentage of additional ingredients in the compositions
of the invention depends on the percentages of Octanoyl Carnosine,
Palmitoyl GHK, and/or GEKG present in a given composition (i.e.,
the additional ingredients make up the balance of the composition).
If carriers (either singularly, such as water, or complex
co-solvents) are used, they may make up the entire balance of the
compositions.
[0336] When preparing compositions combining peptide Octanoyl
Carnosine (or one or more derivatives thereof) with other peptides,
including but not limited to peptide Palmitoyl-GHK (or one or more
derivatives thereof) and/or peptide GEKG (or one or more
derivatives thereof), those peptides can be adding either in their
solid form, or solubilized in a suitable solvent, excipient, and/or
carrier for topical application, at any stage of the composition
preparation process. Often, the peptides can be added as part of
the aqueous phase during the preparation process of the
composition. For instance in case of oil-in-water type
compositions, hydrogels, and aqueous systems, the peptides can be
added as the last step to the previously prepared bulk of the
composition by mixing the peptides, which have been previously
incorporated into a solvent, excipient, and/or carrier for topical
application, to the bulk of the composition at lower temperatures
(e.g., typically below 60.degree. C.; and ideally between about
20.degree. to 40.degree. C.). For example, a solvent, excipient,
and/or carrier suitable to incorporate the peptides into the
composition can include one or more of the following ingredients:
water, glycerin, propylene glycol, propanediol, butylene glycol,
hexylene glycol, low molecular weight PEGs such as PEG-400,
cyclodextrins, ethanol, and/or any mixtures thereof. Additionally,
this solvent, excipient and/or carrier may further contain suitable
surfactants (e.g., Tween 20, Tween 80, etc.) to help solubilize and
stabilize the peptides, and/or anti-microbial preservatives to help
prevent microbial growth and contamination, and/or antioxidants or
iron chelators (e.g., EDTA, etc.) as chemical stabilizers. Water,
glycerin, propylene glycol, propanediol, butylene glycol,
cyclodextrins, and/or any mixtures thereof are the preferred
ingredients.
[0337] However, when preparing compositions combining peptides
which are not or poorly water soluble, or if the peptides cannot be
solubilized as described above using suitable solvents, excipients,
and/or carrier known to the person of skill in the art, the
peptides can be added as part of the oil phase during the
preparation process of the composition. Thereby, the peptides can
be either added to the oil phase in their solid form, or
solubilized in a suitable solvent or carrier system (e.g., oils)
suitable for topical application and able to solubilize or
incorporate the peptides.
[0338] Alternatively, instead of at the end of the preparation
process of an oil-in-water composition for example, Octanoyl
Carnosine (or one or more derivatives thereof) may be added already
at an earlier step in the process. Thereby, Octanoyl Carnosine (or
one or more derivatives thereof) can be added when preparing the
water phase of the composition; either in its solid form, or
solubilized in a suitable solvent or carrier system for Octanoyl
Carnosine (or one or more derivatives thereof).
[0339] When combining Octanoyl Carnosine (or one or more
derivatives thereof) with both Palmitoyl-GHK (or one or more
derivatives thereof) and GEKG (or one or more derivatives thereof),
the optimal weight ratio of Octanoyl Carnosine (or one or more
derivatives thereof) to Palmitoyl-GHK (or one or more derivatives
thereof) to GEKG (or one or more derivatives thereof) is 4 parts
Octanoyl Carnosine (or one or more derivatives thereof): 1 part
Palmitoyl-GHK (or one or more derivatives thereof): 5 parts GEKG
(or one or more derivatives thereof) (parts refer to parts per
weight), as discovered during in vitro studies for hyaluronic acid
formation (see Example 5, infra). Thus, one of the preferred
composition contains Octanoyl Carnosine (or one or more derivatives
thereof), Palmitoyl-GHK (or one or more derivatives thereof) and
GEKG (or one or more derivatives thereof) at a weight ratio of
4:1:5; whereas Octanoyl Carnosine (or one or more derivatives
thereof) is present in the compositions in proportions between
0.001% and 1%, in a carrier or excipient acceptable for topical
application.
[0340] In accordance to the present invention, the following
illustrates suitable compositions for topical applications
containing either 0.01%, 0.013%, 0.02%, or 0.1% of the combination
of Octanoyl Carnosine, Palmitoyl-GHK and GEKG (or one or more
derivatives thereof) at a weight ratio of 4:1:5.
[0341] Furthermore, also in accordance to the present invention,
the following also illustrates suitable compositions for topical
applications containing either 0.01%, 0.1%, 0.25%, 0.5%, 1%, or 5%
of Octanoyl Carnosine (or one or more derivatives thereof).
[0342] Provided herein is information on the preparation of such
exemplary compositions. However, the person of skill in the art
will understand that any other suitable methods can also be used to
prepare compositions in accordance with the instant invention.
[0343] The compositions obtained as described in the following were
tested for physicochemical stability (including, but not limited
to, changes in color, odor, viscosity, pH, and/or appearance) under
accelerated conditions (40.degree. C. to 50.degree. C.) for up to
three to six months. During this period, the compositions were
stable; meaning that the color, odor, viscosity, pH, and the
appearance did not change, or only changed to a limited and
acceptable extent (.+-.10% from baseline). If indicated, testing
for chemical stability of the drug actives (e.g., OTC drug actives
such as sunscreen actives, hydroquinone, retinoic acid, acne drug
active, rosacea drug active, etc.) in the compositions were
additionally realized by acceptable analytical methods (e.g., by
HPLC, etc.).
[0344] The local tolerability (irritant and allergic contact
dermatitis potential) of the compositions provided herein was
determined by patch tests with challenge (repeated insult patch
test) on the back of humans, and, during in use tests for few of
the compositions. Those tests revealed that these compositions are
of acceptable local tolerability. This means that the compositions
intended for cosmetic, feminine, personal care or hygiene use are
neither contact irritants nor contact allergens. The compositions
intended for pharmaceutical use (i.e., medicaments) are of a
negligible to minor potential for inducing irritant and allergic
contact dermatitis in humans. Accordingly, those compositions are
suitable for topical use since they were shown to be stable and of
acceptable local tolerability.
[0345] The compositions can be filled into suitable packaging
(containers) such as, for example, tubes, pumps, airless pumps,
jars, bottles, pens, aerosol containers, or other containers
depending on use and administration. The compositions are generally
commercialized in those containers.
Composition A1: Example of a Serum Containing 0.1% of Octanoyl
Carnosine
[0346] A serum containing 0.1% of Octanoyl Carnosine containing
high levels of hyaluronic acid was prepared as described above and
contained the following other components in order of descending
predominance: WATER, SODIUM HYALURONATE, CITRIC ACID,
PHENOXYETHANOL, SODIUM METHYLPARABEN, SODIUM BUTYLPARABEN, SODIUM
PROPYLPARABEN, and SODIUM CHLORIDE.
Composition A2: Example of a Serum Containing 0.25% of Octanoyl
Carnosine
[0347] A serum containing 0.25% of Octanoyl Carnosine containing
high levels of hyaluronic acid was prepared as described above and
contained the following other components in order of descending
predominance: WATER, SODIUM HYALURONATE, CITRIC ACID,
PHENOXYETHANOL, SODIUM METHYLPARABEN, SODIUM BUTYLPARABEN, SODIUM
PROPYLPARABEN, and SODIUM CHLORIDE.
Composition A3: Example of a Serum Containing 0.5% of Octanoyl
Carnosine
[0348] A serum containing 0.5% of Octanoyl Carnosine containing
high levels of hyaluronic acid was prepared as described above and
contained the following other components in order of descending
predominance: WATER, SODIUM HYALURONATE, CITRIC ACID,
PHENOXYETHANOL, SODIUM METHYLPARABEN, SODIUM BUTYLPARABEN, SODIUM
PROPYLPARABEN, and SODIUM CHLORIDE.
Composition A4: Example of a Serum Containing 1% of Octanoyl
Carnosine
[0349] A serum containing 1% of Octanoyl Carnosine containing high
levels of hyaluronic acid was prepared as described above and
contained the following other components in order of descending
predominance: WATER, SODIUM HYALURONATE, CITRIC ACID,
PHENOXYETHANOL, SODIUM METHYLPARABEN, SODIUM BUTYLPARABEN, SODIUM
PROPYLPARABEN, and SODIUM CHLORIDE.
Composition A5: Example of a Serum Containing 5% of Octanoyl
Carnosine
[0350] A serum containing 5% of Octanoyl Carnosine containing high
levels of hyaluronic acid was prepared as described above and
contained the following other components in order of descending
predominance: WATER, SODIUM HYALURONATE, CITRIC ACID,
PHENOXYETHANOL, SODIUM METHYLPARABEN, SODIUM BUTYLPARABEN, SODIUM
PROPYLPARABEN, and SODIUM CHLORIDE.
Composition B1: Example of Oil-in-Water Cream Containing 0.01% of
Octanoyl Carnosine
[0351] An oil-in-water cream containing 0.01% Octanoyl Carnosine,
free of parabens and triethanolamine, was prepared as described
above and contained the following other components in order of
descending predominance: water, octyldodecanol, decyl oleate,
glyceryl stearate, propanediol, glycerin, stearic acid, Triticum
vulgare (wheat) germ oil, ceteareth-20, cetyl alcohol, Borago
officinalis (borage) seed oil, dimethicone, Triticum vulgare
(wheat) seed extract, myreth-3 myristate, ceteareth-12, tocopheryl
acetate, tocopherol, cetearyl alcohol, cetyl palmitate, carbomer,
disodium EDTA, sodium hydroxide, phenoxyethanol,
ethylhexylglycerin, hexylene glycol, and caprylyl glycol.
Composition B2: Example of Oil-in-Water Cream Containing 0.1% of
Octanoyl Carnosine
[0352] An oil-in-water cream containing 0.1% Octanoyl Carnosine,
free of parabens and triethanolamine, was prepared as described
above and contained the following other components in order of
descending predominance: water, octyldodecanol, decyl oleate,
glyceryl stearate, propanediol, glycerin, stearic acid, Triticum
vulgare (wheat) germ oil, ceteareth-20, cetyl alcohol, Borago
officinalis (borage) seed oil, dimethicone, Triticum vulgare
(wheat) seed extract, myreth-3 myristate, ceteareth-12, tocopheryl
acetate, tocopherol, cetearyl alcohol, cetyl palmitate, carbomer,
disodium EDTA, sodium hydroxide, phenoxyethanol,
ethylhexylglycerin, hexylene glycol, and caprylyl glycol.
Composition B3: Example of Oil-in-Water Cream Containing 0.5% of
Octanoyl Carnosine
[0353] An oil-in-water cream containing 0.5% Octanoyl Carnosine,
free of parabens and triethanolamine, was prepared as described
above and contained the following other components in order of
descending predominance: water, octyldodecanol, decyl oleate,
glyceryl stearate, propanediol, glycerin, stearic acid, Triticum
vulgare (wheat) germ oil, ceteareth-20, cetyl alcohol, Borago
officinalis (borage) seed oil, dimethicone, Triticum vulgare
(wheat) seed extract, myreth-3 myristate, ceteareth-12, tocopheryl
acetate, tocopherol, cetearyl alcohol, cetyl palmitate, carbomer,
disodium EDTA, sodium hydroxide, phenoxyethanol,
ethylhexylglycerin, hexylene glycol, and caprylyl glycol.
Composition B4: Example of Oil-in-Water Cream Containing 1% of
Octanoyl Carnosine
[0354] An oil-in-water cream containing 1% Octanoyl Carnosine, free
of parabens and triethanolamine, was prepared as described above
and contained the following other components in order of descending
predominance: water, octyldodecanol, decyl oleate, glyceryl
stearate, propanediol, glycerin, stearic acid, Triticum vulgare
(wheat) germ oil, ceteareth-20, cetyl alcohol, Borago officinalis
(borage) seed oil, dimethicone, Triticum vulgare (wheat) seed
extract, myreth-3 myristate, ceteareth-12, tocopheryl acetate,
tocopherol, cetearyl alcohol, cetyl palmitate, carbomer, disodium
EDTA, sodium hydroxide, phenoxyethanol, ethylhexylglycerin,
hexylene glycol, and caprylyl glycol.
Composition B5: Example of Oil-in-Water Cream Containing 5% of
Octanoyl Carnosine
[0355] An oil-in-water cream containing 5% Octanoyl Carnosine, free
of parabens and triethanolamine, was prepared as described above
and contained the following other components in order of descending
predominance: water, octyldodecanol, decyl oleate, glyceryl
stearate, propanediol, glycerin, stearic acid, Triticum vulgare
(wheat) germ oil, ceteareth-20, cetyl alcohol, Borago officinalis
(borage) seed oil, dimethicone, Triticum vulgare (wheat) seed
extract, myreth-3 myristate, ceteareth-12, tocopheryl acetate,
tocopherol, cetearyl alcohol, cetyl palmitate, carbomer, disodium
EDTA, sodium hydroxide, phenoxyethanol, ethylhexylglycerin,
hexylene glycol, and caprylyl glycol.
Composition C: Example of Oil-in-Water Cream Containing 0.01% Total
Peptides
[0356] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
was prepared as described above and contained the following other
components in order of descending predominance: water, hydrogenated
peanut oil, glycerin, cetearyl ethylhexanoate, cetearyl alcohol,
PEG-8 C12-18 alkyl ester, PPG-25-laureth-25, PEG-5 pentaerythrityl
ether, hydroxyethylcellulose, cetyl alcohol, cetyl palmitate,
glyceryl stearate, sodium chloride, ascorbyl palmitate, glucose,
simethicone, tocopheryl acetate, citric acid, ricinoleth-40,
potassium chloride and magnesium chloride. The cream further
contained a mixture of metylparaben, propylparaben and
imidazolidinyl urea for antimicrobial preservation.
Composition D1: Example of Oil-in-Water Cream Containing 0.01%
Total Peptides
[0357] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
was prepared as described above and contained the following other
components in order of descending predominance: water,
octyldodecanol, glyceryl stearate decyl oleate, glycerin, propylene
glycol, triticum vulgare (wheat germ oil), stearic acid, cetyl
alcohol, ceteareth 20, myreth-3 myristate, ceteareth 12, cetearyl
alcohol, cetyl palmitate, tocopheryl acetate, dimethicone, borago
officinalis (borage seed oil), carbomer, triethanolamine,
methylparaben, propylparaben, glycosphingolipids (e.g., from
Triticum vulgare (wheat) seed extract), disodium EDTA and BHT.
[0358] The cream further contained a mixture of phenoxyethanol,
ethylparaben, butylparaben, methyl-isothiazolinone and
methylchloroisothiazolinone for antimicrobial preservation.
Composition D2: Example of Oil-in-Water Cream Containing 0.01%
Total Peptides
[0359] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
was prepared as described above and contained the following other
components in order of descending predominance: water,
octyldodecanol, glyceryl stearate decyl oleate, glycerin, propylene
glycol, triticum vulgare (wheat germ oil), stearic acid, cetyl
alcohol, ceteareth 20, myreth-3 myristate, ceteareth 12, cetearyl
alcohol, cetyl palmitate, tocopheryl acetate, dimethicone, borago
officinalis (borage seed oil), carbomer, triethanolamine,
methylparaben, sodium methylparaben, propylparaben, ethylparaben,
butylparaben, glycosphingolipids (e.g., from Triticum vulgare
(wheat) seed extract), disodium EDTA and BHT.
Composition E: Example of Oil-in-Water Cream Containing 0.01% Total
Peptides
[0360] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
was prepared as described above and contained the following other
components in order of descending predominance: water,
octyldodecanol, glyceryl stearate decyl oleate, glycerin, propylene
glycol, triticum vulgare (wheat germ oil), stearic acid, cetyl
alcohol, ceteareth 20, myreth-3 myristate, ceteareth 12, cetearyl
alcohol, cetyl palmitate, tocopheryl acetate, dimethicone, borago
officinalis (borage seed oil), carbomer, methylparaben,
propylparaben, glycosphingolipids (e.g., from Triticum vulgare
(wheat) seed extract), and disodium EDTA. The cream further
contained tocopherol, sodium hydroxide, and a mixture of
phenoxyethanol, ethylparaben, butylparaben for antimicrobial
preservation.
Composition F: Example of Oil-in-Water Cream Containing 0.013%
Total Peptides
[0361] An oil-in-water cream containing 0.013% of the combination
of Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio
4:1:5 was prepared as described above and contained the following
other components in order of descending predominance: water,
caprylic/capric triglyceride, C12-20 acid PEG-8 ester, butylene
glycol, glycerin, saccharide isomerate, PEG-8, cetyl alcohol,
caprylyl glycol, potassium cetyl phosphate, carbomer, bisabolol,
ascorbyl tetraisopalmitate, caffeine, disodium EDTA, phospholipids,
glycyrrhetinic acid, sodium hyaluronate, sodium polyacrylate,
citric acid, propylparaben, tocopherol, beech tree bud extract
(fagus sylvatica extract), palm oil (elaeis guineensis),
tocotrienols, ascorbyl palmitate, squalene, ascorbic acid and
phytosterols. The cream further contained a mixture of
phenoxyethanol, methylparaben, butylparaben, and ethylparaben for
antimicrobial preservation.
Composition G: Example of Oil-in-Water Cream Containing 0.013%
Total Peptides
[0362] An oil-in-water cream containing 0.013% of the combination
of Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio
4:1:5 was prepared as described above and contained the following
other components in order of descending predominance: WATER, C12-20
ACID PEG-8 ESTER, PETROLATUM, CAPRYLIC/CAPRIC TRIGLYCERIDE,
HYDROGENATED POLYISOBUTENE, GLYCERIN, SACCHARIDE ISOMERATE,
HYDROXYETHYL ACRYLATE/SODIUM ACRYLOYLDIMETHYL TAURATE COPOLYMER,
BUTYLENE GLYCOL, ETHYLHEXYLGLYCERIN, ISOHEXADECANE, POTASSIUM CETYL
PHOSPHATE, DIOSCOREA VILLOSA (WILD YAM) ROOT EXTRACT, BISABOLOL,
ASCORBYL TETRAISOPALMITATE, CAFFEINE, DISODIUM EDTA, PHOSPHOLIPIDS,
GLYCYRRHETINIC ACID, SODIUM HYALURONATE, ACRYLATES/C10-30 ALKYL
ACRYLATE CROSSPOLYMER, CITRIC ACID, SODIUM CITRATE, SODIUM
HYDROXIDE, TOCOPHEROL, BEECH TREE BUD EXTRACT (FAGUS SYLVATICA
EXTRACT), PALM OIL (ELAEIS GUINEENSIS), TOCOTRIENOLS, SQUALENE,
PHYTOSTEROLS, PHENOXYETHANOL, CHLORPHENESIN, POLYSORBATE 60, BENZYL
ALCOHOL, BENZOIC ACID
Composition H: Example of Oil-in-Water Cream Containing 0.01% Total
Peptides
[0363] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
was prepared as described above and contained the following other
components in order of descending predominance: water,
caprylic/capric triglyceride, C12-20 acid PEG-8 ester,
coco-caprylate/caprate, butylene glycol, dimethicone, phenyl
trimethicone, biosaccharide gum-1, glycerin, cetyl alcohol,
phenoxyethanol, saccharide isomerate, carbomer, potassium cetyl
phosphate, borago officinalis (borage seed oil), ascorbyl
tetraisopalmitate, caprylyl glycol, methylparaben, disodium EDTA,
chondrus crispus (carrageenan), sodium hyaluronate, elaeis
guineensis (palm) oil, tocotrienols, phytosterols, butylparaben,
ethylparaben, PEG-8, isobutylparaben, propylparaben, tocopherol,
citric acid, ascorbyl palmitate, squalene and ascorbic acid.
Composition I: Example of Oil-in-Water Cream Containing 0.01% Total
Peptides
[0364] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
was prepared as described above and contained the following other
components in order of descending predominance: water, ethylhexyl
methoxycinnamate, C12-20 acid PEG-8 ester, caprylic/capric
triglyceride, coco-caprylate/caprate, butylene glycol, butyl
methoxydibenzoylmethane, cetyl alcohol, biosaccharide gum-1,
glycerin, C12-15 alkyl benzoate, saccharide isomerate,
phenoxyethanol, caprylyl glycol, titanium dioxide, potassium cetyl
phosphate, carbomer, borago officinalis seed oil, ascorbyl
tetraisopalmitate, methylparaben, sodium hydroxide, disodium EDTA,
chondrus crispus (carrageenan), sodium hyaluronate, elaeis
guineensis (palm) oil, tocotrienols, phytosterols, butylparaben,
aluminum stearate, polyhydroxystearic acid, ethylparaben, alumina,
PEG-8, isobutylparaben, propylparaben, tocopherol, citric acid,
BHT, ascorbyl palmitate, squalene and ascorbic acid.
Composition J: Example of Oil-in-Water Cream Containing 0.01% Total
Peptides
[0365] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
which provides additionally sun protection (e.g., of about SPF30)
properties due to the presence of sunscreen actives including
octinoxate and zinc oxide was prepared as described above and
contained the following other components: sunscreens actives:
octinoxate (about 7.5%) and zinc oxide (about 7.3%). Remaining
ingredients in order of descending predominance: Water,
Caprylic/Capric Triglyceride, Hydrogenated C6-14 Olefin Polymers,
Hexyldecanol, Glycerin, Glyceryl Stearate, PEG-100 Stearate,
Potassium Cetyl Phosphate, Hydrogenated Palm Glycerides,
Microcrystalline Cellulose, Camelia Sinensis Extract, Silica,
Sodium Ascorbyl Phosphate, Tocopheryl Acetate, Squalane,
Hydroxyethyl Acrylate/Sodium Acryloyldimethyl Taurate Copolymer,
Polyhydroxystearic Acid, Steareth-21, Melanin, Cetearyl Alcohol,
Sodium Hyaluronate, Polysorbate 60, Triethoxycaprylsilane, Disodium
EDTA, Xanthan Gum, Styrene/Acrylates Copolymer, Phenoxyethanol,
Methylparaben, Ethylparaben, Propylparaben.
Composition K: Example of Oil-in-Water Cream Containing 0.01% Total
Peptides
[0366] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5,
free of parabens, triethanolamine and BHT, was prepared as
described above and contained the following other components in
order of descending predominance: water, octyldodecanol, decyl
oleate, glyceryl stearate, propanediol, glycerin, stearic acid,
Triticum vulgare (wheat) germ oil, ceteareth-20, cetyl alcohol,
Borago officinalis (borage) seed oil, dimethicone, Triticum vulgare
(wheat) seed extract, myreth-3 myristate, ceteareth-12, tocopheryl
acetate, tocopherol, cetearyl alcohol, cetyl palmitate, carbomer,
disodium EDTA, sodium hydroxide, phenoxyethanol,
ethylhexylglycerin, hexylene glycol, and caprylyl glycol.
Composition L: Example of Oil-in-Water Serum Containing 0.02% Total
Peptides (Excluding TRIFLUORACETYL TRIPEPTIDE-2 which is Present in
this Composition as an Additional Peptide)
[0367] An oil-in-water serum containing 0.02% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
was prepared as described above and contained the following other
components in order of descending predominance: WATER, GLYCERIN,
CAPRYLIC/CAPRIC TRIGLYCERIDE, HYDROXYETHYL ACRYLATE/SODIUM
ACRYLOYLDIMETHYL TAURATE COPOLYMER, PULLULAN, SQUALANE, AMINOPROPYL
ASCORBYL PHOSPHATE, BUTYLENE GLYCOL, SODIUM HYALURONATE, ACETYL
GLUCOSAMINE, ARGININE, PROLINE, GLYCINE, GLUTAMINE, TRIFLUORACETYL
TRIPEPTIDE-2, ALGAE EXTRACT, POLYSORBATE 60, DEXTRAN,
ETHYLHEXYLGLYCERIN, ETHYLENE/ACRYLIC ACID COPOLYMER, DISODIUM EDTA,
SODIUM HYDROXIDE, CHLORPHENESIN, PHENOXYETHANOL
Composition M: Example of Oil-in-Water Serum Containing 0.1% Total
Peptides (Excluding TRIFLUORACETYL TRIPEPTIDE-2 which is Present in
this Composition as an Additional Peptide)
[0368] An oil-in-water serum containing 0.1% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
was prepared as described above and contained the following other
components in order of descending predominance: WATER, GLYCERIN,
CAPRYLIC/CAPRIC TRIGLYCERIDE, HYDROXYETHYL ACRYLATE/SODIUM
ACRYLOYLDIMETHYL TAURATE COPOLYMER, PULLULAN, SQUALANE, AMINOPROPYL
ASCORBYL PHOSPHATE, BUTYLENE GLYCOL, SODIUM HYALURONATE, ACETYL
GLUCOSAMINE, ARGININE, PROLINE, GLYCINE, GLUTAMINE, TRIFLUORACETYL
TRIPEPTIDE-2, ALGAE EXTRACT, POLYSORBATE 60, DEXTRAN,
ETHYLHEXYLGLYCERIN, ETHYLENE/ACRYLIC ACID COPOLYMER, DISODIUM EDTA,
SODIUM HYDROXIDE, CHLORPHENESIN, PHENOXYETHANOL
Composition N: Example of Oil-in-Water Lotion Containing 0.01%
Total Peptides
[0369] An oil-in-water lotion containing 0.01% of the combination
of Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio
4:1:5 which provides additional anti-properties due to the presence
of retinol (e.g., embedded in microcapsules for controlled release)
was prepared as described above and contained the following other
components in order of descending predominance: Water,
Caprylic/Capric Triglyceride, Glycerin, Cetearyl Alcohol, C10-30
Cholesterol/Lansterol Esters, Cetyl Ricinoleate, Cetyl Alcohol,
Dimethicone, Polysorbate 60, Methyl Methacrylate/Glycol
Dimethacrylate Crosspolymer, Benzyl Alcohol, Retinol, Ascorbic
Acid, Ascorbyl Palmitate, Bisabolol, Tocopheryl Acetate,
Cyclopentasiloxane, Cyclohexasiloxane, PEG-10 Soy Sterol, Stearic
Acid, BHT, Propyl Gallate, Disodium EDTA, Magnesium Aluminum
Silicate, Phenoxyethanol, Polysorbate 20, Triethanolamine, and
Methylparaben.
Composition O1: Example of a Serum Containing 0.01% Total
Peptides
[0370] An serum containing 0.01% of the combination of Octanoyl
Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5 and
containing high levels of hyaluronic acid was prepared as described
above and contained the following other components in order of
descending predominance: WATER, SODIUM HYALURONATE, CITRIC ACID,
PHENOXYETHANOL, SODIUM METHYLPARABEN, SODIUM BUTYLPARABEN, SODIUM
PROPYLPARABEN, and SODIUM CHLORIDE.
Composition O2: Example of a Serum Containing 0.1% Total
Peptides
[0371] An serum containing 0.1% of the combination of Octanoyl
Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5 and
containing high levels of hyaluronic acid was prepared as described
above and contained the following other components in order of
descending predominance: WATER, SODIUM HYALURONATE, CITRIC ACID,
PHENOXYETHANOL, SODIUM METHYLPARABEN, SODIUM BUTYLPARABEN, SODIUM
PROPYLPARABEN, and SODIUM CHLORIDE.
Composition P: Example of a Gel Containing 0.01% Total Peptides
[0372] A gel (e.g., hydrogel) containing 0.01% of the combination
of Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio
4:1:5 was prepared as described above and contained the following
other components in order of descending predominance: WATER (AQUA),
BUTYLENE GLYCOL, GLYCERIN, CARBOMER, PEG-8, CAPRYLYL GLYCOL,
CARRAGEENAN (CHONDRUS CRISPUS), PHENOXYETHANOL, DISODIUM EDTA,
METHYLPARABEN, BUTYLPARABEN, SODIUM HYALURONATE, ETHYLPARABEN,
SODIUM POLYACRYLATE, ISOBUTYLPARABEN, and PROPYLPARABEN.
Composition Q: Example of Oil-in-Water Cream Containing 0.01% Total
Peptides
[0373] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
which provides additionally skin lightening properties due to the
presence of skin lighting agents (as disclosed in WO 2010/083368)
was prepared as described above and contained the following other
components:
TABLE-US-00001 INGREDIENT INCI % BY NO. PHASE (TRADE NAME)
DESIGNATION SUPPLIER WEIGHT 1 A DEIONIZED WATER WATER (AQUA) 63.30
2 A NA.sub.2EDTA DISODIUM EDTA AKZO/ 0.100 DEWOLF 3 A KELTROL CG-T
XANTHAN GUM CP KELCO/ 0.300 UNIVAR 4 B LIPOWAX D CETEARYL LIPO
8.250 ALCOHOL CETEARETH-20 5 B LIPO GMS 450 GLYCERYL LIPO 6.000
STEARATE 6 B CERAPHYL 230 DIISOPROPYL ISP SUTTON 5.000 ADIPATE 7 B
DC TORAY FZ-3196 CAPRYLYL DOW 3.000 METHICONE CORNING/ UNIVAR 8 B
DC 200 FLUID 100 DIMETHICONE DOW 1.000 CST CORNING/ UNIVAR 9 B
LIPOVOL J SIMMONDSIA LIPO 1.000 CHINENSIS (JOJOBA) SEED OIL 10 B
SHEA BUTTER HMP BUTYROSPERMUM EARTH 1.000 PARKII (SHEA SUPPLIED
BUTTER) PRODUCTS 11 B VITAMIN E ACETATE DL-ALPHA BASF/ 0.200 OIL
TOCOPHERYL CHEMCENTRAL (USP, FCC) ACETATE 12 C DEIONIZED WATER
WATER (AQUA) 0.100 13 C ELESTAB CPN CHLORPHENESIN COGNIS 0.300
ULTRA PURE 14 C PHENOXETOL PHENOXYETHANOL CLARIANT 0.600 15 C
SEPIWHITE MSH UNDECYLENOYL SEPPIC 0.500 PHENYLALANINE 16 C SODIUM
SODIUM DR. PAUL 3.000 GLYCEROPHOSPHATE GLYCEROPHOSPHATE LOHMANN
(Ph. Eur. 6 Ed, Item# 500012045500) 17 C L-LEUCINE LEUCINE
AJINOMOTO 1.000 18 C1 CITRIC ACID 50% CITRIC ACID PCI 1.920
SOLUTION (TO pH 4.5-5.0) 19 C2 GLYCERIN 99.7% GLYCERIN ACME- 2.000
(USP) HARDESTY 20 C2 SYMWHITE 377 PHENYLETHYL KAH/SYMRISE 0.500
RESORCINOL 21 C2 VITAGEN AMINOPROPYL BASF 0.500 ASCORBYL PHOSPHATE
22 D SIMULGEL INS 100 HYDROXYETHYL SEPPIC 0.420 ACRYLATE/SODIUM
ACRYLOYLDIMETHYL TAURATE COPOLYMER ISOHEXADECANE POLYSORBATE 60
TOTAL 99.99
[0374] Such compositions were generally prepared in a clean and
sanitized stainless steel vessel as described herein below: [0375]
PHASE A: DISPERSE KELTROL IN WATER, MIX UNTIL ALL HYDRATES; [0376]
ADD REMAINING PHASE A INGREDIENTS, HEAT TO ABOUT 75.degree. C.
WHILE MIX UNTIL ALL DISSOLVES. [0377] PHASE B: COMBINE PHASE B
INGREDIENTS IN A SEPARATE VESSEL AND MIX WHILE HEATING TO
75.degree. C.; ONCE ALL WAXES MELT AND PHASE IS AT TEMP AND
UNIFORM, SLOWLY ADD TO PHASE A; COOL TO 35.degree. C. [0378] PHASE
C: COMBINE PHASE C INGREDIENTS WITH MECHANICAL STIRRING UNIT AND
MIX WITH MODERATE AGITATION [0379] PHASE C1: USE PHASE C1 TO ADJUST
pH OF PHASE C TO 4.0-4.5 [0380] PHASE C2: COMBINE PHASE C2 AND MIX
WHILE HEATING SLIGHTLY TO 40.degree. C.; CONTINUE MIXING UNTIL
POWDERS DISSOLVE THEN ADD TO PHASE C; ADD PHASE C TO BATCH WITH
MODERATE AGITATION [0381] PHASE D: ADD PHASE D TO BATCH, MIX UNTIL
UNIFORM; [0382] HOMOGENIZE THE BATCH AT 3500 RPM FOR 5 MINUTES;
SWITCH TO IMPELLER MIXING; COOL TO ROOM TEMPERATURE.
Composition R: Example of Oil-in-Water Cream Containing 0.01% Total
Peptides
[0383] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
which provides additionally skin lightening properties due to the
presence of skin lighting agents including hydroquinone (as
disclosed in WO 2010/083368) was prepared as described above and
contained the following other components:
TABLE-US-00002 INGREDIENT INCI % BY NO. PHASE (TRADE NAME)
DESIGNATION SUPPLIER WEIGHT 1 A DEIONIZED WATER WATER (AQUA) 59.670
2 A NA.sub.2EDTA DISODIUM EDTA AKZO 0.100 3 A KELTROL CG-T XANTHAN
GUM CP KELCO 0.300 4 A ELESTAB CPN CHLORPHENESIN COGNIS 0.300 ULTRA
PURE 5 A PHENOXETOL PHENOXYETHANOL CLARIANT 0.600 6 A SEPIWHITE MSH
UNDECYLENOYL SEPPIC 0.500 PHENYLALANINE 7 A SODIUM SODIUM DR. PAUL
3.000 GLYCEROPHOSPHATE GLYCEROPHOSPHATE LOHMANN (Ph. Eur. 6 Ed,
Item# 500012045500) 8 A L-LEUCINE LEUCINE AJINOMOTO 1.000 9 B
LIPOWAX D CETEARYL LIPO 6.000 ALCOHOL CETEARETH-20 10 B LIPO GMS
450 GLYCERYL LIPO 6.000 STEARATE 11 B CERAPHYL 230 DIISOPROPYL ISP
SUTTON 3.000 ADIPATE 12 B DC TORAY FZ-3196 CAPRYLYL DOW 3.000
METHICONE CORNING 13 B DC 200 FLUID 100 DIMETHICONE DOW 1.000 CST
CORNING/UNIVAR 14 B LIPOVOL J SIMMONDSIA LIPO 1.000 CHINENSIS
(JOJOBA) SEED OIL 15 B SHEA BUTTER HMP BUTYROSPERMUM EARTH 1.000
PARKII (SHEA SUPPLIED BUTTER) PRODUCTS 16 B VITAMIN E DL-ALPHA
BASF/ 0.200 ACETATE OIL TOCOPHERYL CHEMCENTRAL (USP, FCC) ACETATE
17 C CITRIC ACID 50% CITRIC ACID PCI 1.920 SOLUTION (TO pH 4.5-5.0)
18 D EASTMAN .TM. HYDROQUINONE EASTMAN/ 4.000 HYDROQUINONE
CHEMPOINT (USP GRADE) 19 E SODIUM SODIUM UPI 0.400 METABISULFITE
METABISULFITE (NF/FCC) 20 F GLYCERIN 99.7% GLYCERIN ACME- 2.000
(USP) HARDESTY 21 F SYMWHITE 377 PHENYLETHYL KAH/SYMRISE 0.500
RESORCINOL 22 F VITAGEN AMINOPROPYL BASF 0.500 ASCORBYL PHOSPHATE
23 G SIMULGEL INS 100 HYDROXYETHYL SEPPIC 4.000 ACRYLATE/SODIUM
ACRYLOYLDIMETHYL TAURATE COPOLYMER ISOHEXADECANE POLYSORBATE 60
TOTAL 99.99
[0384] Such compositions were generally prepared in a clean and
sanitized stainless steel vessel, which was suitable for blending
products containing hydroquinone, as described herein below: [0385]
PHASE A: DISPERSE KELTROL IN WATER, MIX UNTIL ALL HYDRATES; ADD
EDTA, MIX UNTIL ALL DISSOLOVES; [0386] ADD REMAINING PHASE A
INGREDIENTS, HEAT TO 75.degree. C. WHILE MIX UNTIL ALL DISSOLVES.
[0387] PHASE B: COMBINE PHASE B INGREDIENTS, HEAT TO 75.degree. C.,
MIX UNTIL ALL MELTED AND UNIFORM; [0388] WHEN BOTH PHASE A AND
PHASE B AT 75.degree. C., ADD PHASE B INTO PHASE A WITH AGITATION
MIX FOR 10 MINUTES, START COOLING TO 50.degree. C. [0389] PHASE C:
ADJUST pH WITH PHASE C TO pH 4.5-5.0, COOL TO 45.degree. C. [0390]
PHASE D: ADD PHASE D TO BATCH WITH MIX, MIX UNTIL ALL DISSOLVES AND
UNIFORM. [0391] PHASE E: ADD PHASE E TO BATCH WITH MIXING, MIX
UNTIL ALL DISSOLVES. [0392] PHASE F: COMBINE PHASE F INGREDIENTS,
SLIGHTLY HEAT AND MIX UNTIL ALL DISSOLVES, ADD TO THE BATCH. [0393]
PHASE G: ADD PHASE G TO BATCH, MIX UNTIL UNIFORM; [0394] HOMOGENIZE
THE BATCH AT 3500 RPM FOR 5 MINUTES, SWITCH TO IMPELLER MIXER, MIX;
[0395] ADJUST pH WITH PHASE C TO pH 4.5-5.0 IF NECESSARY.
Composition S: Example of Oil-in-Water Serum Containing 0.01% Total
Peptides
[0396] An oil-in-water serum containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
which provides additionally anti-rosacea or skin redness reducing
properties due to the presence of antimicrobial peptide
sequestering agents such as the sodium salt of dextran sulfate (as
disclosed in WO 2011/109469) was prepared as described above and
contained the following other components:
TABLE-US-00003 Ingredient % by weight Phase (Trade Name) INCI Name
Supplier(s) (% w) A Water Water (Aqua) 60.03 A Na2EDTA Disodium
EDTA Akzo/DeWolf 0.1 A Keltrol CG-SFT Xanthan Gum CP Kelco/ 0.25
Univar A Structure XL Hydroxypropyl Starch National Starch 1.0
Phosphate A Simulgel INS 100 Hydroxyethyl Seppic 1.0
Acrylate/Sodium Acryloyldimethyl Taurate Copolymer (and)
Isohexadecane (and) Polysorbate-60 B Water 15 B Oristract CF
Caffeine Orient Stars 1.5 B Dextran Sulfate Dextran Sodium Sulfate
MP Biomedical/ 0.5 Sodium Salt (av. Spectrum M.W. about 8000) B
Ajidew ZN-100 Zinc PCA Ajinomoto 1 B1 Elestab CPN Ultra
Chlorphenesin Cognis 0.2 Pure B1 Glycerin 99.7% Glycerin
Acme-Hardesty 15 B1 Phenoxetol Phenoxyethanol Clariant/ 0.5
Essential Ingredients C Structure XL Hydroxypropyl Starch National
Starch 1.5 Phosphate C Simulgel INS 100 Hydroxyethyl Seppic 0.41
Acrylate/Sodium Acryloyldimethyl Taurate Copolymer (and)
Isohexadecane (and) Polysorbate-60 D DC Toray FZ- Caprylyl
Methicone Dow Corning/ 2 3196 Univar
[0397] Such compositions were generally prepared in a clean and
sanitized stainless steel vessel as described herein below:
Phase A: Dissolve Na2EDTA into agitating Phase A water. Mix until
uniform. Sprinkle Keltrol slowly into batch. Mix until fully
hydrated. Sprinkle Structure XL into agitating Phase A water. Mix
until fully dispersed, and homogenize at 3500 RPM for 5-6 minutes.
Add Simulgel INS 100 and mix until uniform, then homogenize for
about 4 minutes at 3500 RPM. Phase B: Combine Phase B ingredients,
one by one in a separate vessel while heating to 50-53.degree.
Celsius (not higher than 55.degree. Celsius). Phase B1: In a
separate vessel combine Phase B1 ingredients and heat to 40.degree.
Celsius. Mix until powder is dispersed. Add Phase B1 to Phase B and
mix until clear. Cool to 30.degree. Celsius and add combined Phase
B/B1 to batch. Mix until uniform. Phase C: Add Phase C to batch one
by one to raise viscosity, homogenize after adding the Structure X1
and again after adding the Simulgel INS 100. Phase D: Add Phase D
ingredients to batch and mix until uniform.
Example 5
Results of In Vitro Tests on Examples According to the
Invention
In Vitro Test 1: Stimulation of Gene Expression of Extracellular
Matrix Components Including but not Limited to Collagens I and III
by Octanoyl Carnosine
[0398] Normal human dermal fibroblasts (NHDF); used at the 8.sup.th
passage, were cultured in culture medium (DMEM supplemented with
10% fetal calf serum, 2 mM L-glutamine, 50 U/ml penicillin, and 50
.mu.g/ml streptomycin) in 24-well plates for 48 h at 37.degree. C.
and 5% CO.sub.2 with a renewal of culture medium after 24 h. The
culture medium was then removed, replaced by assay medium (DMEM
culture medium supplemented with 1% fetal calf serum, 2 mM
L-glutamine, 50 U/ml penicillin, and 50 .mu.g/ml streptomycin) and
the cells were further incubated for 24 h. The cells were then
treated with the test peptides, or not (control), or with the
reference TGF-beta 1 and incubated for 24 h. All experimental
conditions were performed 4-times (n=4). At the end of the
incubation, the cells were washed in PBS solution and immediately
frozen at about -80.degree. C. The expression of the markers was
analyzed using quantitative real time polymerase chain reaction
(RT-qPCR) method on mRNA extracted from the cell monolayers for
each treatment; whereby the replicates were pooled before RNA
extraction. Analysis of gene expression was performed in duplicate
(n=2) using a PCR array. Total RNA was extracted from each sample
using TriPure Isolation Reagent.RTM. according to the supplier's
instructions. The amount and quality of RNA were evaluated using a
lab-on-a-chip Bioanalyzer (Agilent Technologies). Potential
contaminant traces of genomic DNA were removed using the DNAfree
system (Ambion). The reverse-transcription of mRNA was conducted in
presence of oligo(dT) and Superscript II reverse-transcriptase.
Quantification of cDNA was performed using Nanovue (GE Healthcare)
and cDNA was adjusted.
[0399] The PCRs were performed using the LightCycler.RTM. system
(Roche Molecular System Inc.) according to the supplier's
instructions. This system allowed rapid and powerful PCRs, after
determining analysis conditions of the test primers. The reaction
mix (10 .mu.l final) was added as follows: 2.5 .mu.l of cDNA,
primers forward and reverse; and reagent mix containing taq DNA
polymerase, SYBR Green I, and MgCl.sub.2.
[0400] The incorporation of the fluorescence in amplified DNA was
continuously measured during the PCR cycles. This resulted in a
"fluorescence intensity" versus "PCR cycle" plot allowing the
evaluation of a relative expression (RE) value for each marker. The
value selected for RE calculations is the "output point" (Ct) of
the fluorescence curve. For a considered marker, the highest is the
cycle number; the lowest is the mRNA quantity. The RE value was
expressed in arbitrary units (AU) according to the formula:
(1/2.sup.number of cycles).times.10.sup.6.
[0401] This evaluation was realized with non-cytotoxic
concentrations of the test peptides. Cytotoxicity of the test
peptides in the assay medium was previously determined in NHDF by
the MTT reduction assay and morphological observations with help of
microscope after a 24 h incubation time.
[0402] This test surprisingly revealed that the peptide Octanoyl
Carnosine is able to significantly stimulate extracellular matrix
components other than only collagen I. It was discovered that
Octanoyl Carnosine at 300 ppm (0.03%) stimulates the formation of
collagen I (alpha 1) by 38%, collagen III (alpha 1) by 120%,
collagen V (alpha 1) by 35%, collagen VI (alpha 1) by 46%, collagen
VII (alpha 1) by 97%, collagen XVI (alpha 1) by 91%, elastin by
51%, laminin (alpha 4) by 20%, hyaluronan synthase 2 by 23%,
fibrillin 1 by 23%, and heparan sulfate protoglycan 2 by 79%.
[0403] Even more unexpectedly, this test surprisingly revealed that
the peptide Octanoyl Carnosine is able to stimulate collagen III
significantly more than collagen I; collagen III (alpha 1)
formation was stimulated by 120%, whereas collagen I (alpha 1) only
by 38%. This property of Octanoyl Carnosine was unexpected as other
peptides which stimulate formation of both collagen I and collagen
III generally stimulate collagen I significantly more than collagen
III. For example, as also demonstrated in this test,
beta-alanyl-histidine (the natural carnosine) at 300 ppm (0.03%)
stimulates formation of collagen III only by 24% but stimulates
collagen I at 28%. Furthermore, TGF-beta 1 (10 ng/ml) was shown to
stimulate formation of collagen III by 164% but at the same time
stimulates collagen I by 245%.
[0404] For the interpretation of the data related to the
stimulation of the formation of extracellular matrix components as
determined during the in vitro experiments, a "significant"
stimulation of the formation of extracellular matrix components
means a stimulation of at least 20% more than measured using a
control material under identical experimental conditions. The
control material is the identical material (here: assay medium) as
used when testing the active (here: peptide(s)) but without the
active(s).
[0405] Consequently, compositions containing Octanoyl Carnosine (or
one or more derivatives thereof) according to the present invention
have therefore great potential in the field of cosmetics,
dermatology, wound healing, and any other areas in need of
treatments for conditions, disorders and diseases where
extracellular matrix components (including, but not limited to,
collagen III) are altered. In addition, such compositions can be
used for maintaining healthy skin, skin rejuvenation, scarless
wound healing, restoration of damaged skin and mucosa, as well as
for the treatment of atrophy of any human tissue including, but not
limited to, vulvovaginal atrophy.
[0406] In contrast, the prior art discloses other peptides
significantly stimulating both collagen I and collagen III
formation, but they stimulate collagen I more than collagen
III.
In Vitro Test 2: Stimulation of Hyaluronic Acid--Combination of
Octanoyl Carnosine with Other Peptides
[0407] Normal human dermal fibroblasts (NHDF); used at the 8.sup.th
passage, were cultured in culture medium (DMEM supplemented with
10% fetal calf serum, 2 mM L-glutamine, 50 U/ml penicillin, and 50
.mu.g/ml streptomycin) in 96-well plates for 24 h at 37.degree. C.
and 5% CO.sub.2.
[0408] Afterwards, the culture medium was removed and replaced by
assay medium (DMEM culture medium supplemented with 2% fetal calf
serum, 2 mM L-glutamine, 50 U/ml penicillin, and 50 .mu.g/ml
streptomycin) containing the test peptides or not (control). The
cells were then incubated for 72 h. All experimental conditions
were performed 3-times (n=3). At the end of the incubation, culture
supernatants were collected and immediately frozen at about
-80.degree. C.
[0409] Hyaluronic acid was quantified in the culture supernatant
using the Duoset Hylaluronan ELISA kit (R&D Systems Ref.
DY3614, 0.37 ng/ml as low detection limit) according to the
provider's specifications.
[0410] This evaluation was realized with non-cytotoxic
concentrations of the test peptides. Cytotoxicity of the test
peptides in the assay medium was previously determined in NHDF by
the MTT reduction assay and morphological observations with help of
microscope after a 72 h incubation time.
[0411] This test surprisingly revealed that the combination of
Octanoyl Carnosine with both Palmitoyl-GHK and GEKG at a weight
ratio of 4 parts Octanoyl Carnosine, 1 part Palmitoyl-GHK, and 5
parts GEKG stimulates synthesis of hyaluronic acid significantly
more than at other ratios. We discovered that a combination of 100
ppm (0.01%) Octanoyl Carnosine, 25 ppm (0.0025%) Palmitoyl-GHK and
125 ppm (0.0125%) GEKG significantly stimulates the synthesis of
hyaluronic acid by 64%.
[0412] This significant stimulation of the synthesis of hyaluronic
acid using Octanoyl Carnosine when combined with Palmitoyl-GHK and
GEKG at a weight ratio of 4:1:5 was unexpected, as other weight
ratios did not produce similar stimulation of hyaluronic acid
synthesis.
[0413] For instance, this test revealed that a combination at the
same total concentration of peptides (250 ppm) at a weight ratio of
1:2:2 (Octanoyl Carnosine:Palmitoyl-GHK:GEKG) only
non-significantly stimulated the synthesis of hyaluronic acid by
8%. Likewise, combinations at a weight ratio of 1:1:2 (tested at
total peptide concentrations of 132 ppm and 400 ppm), 1:3:1 (166
and 500 ppm), and 1:30:1 (320 ppm) non-significantly stimulated the
synthesis of hyaluronic acid by less than 20%.
[0414] Even more surprisingly, this test also revealed that the
combination of Octanoyl Carnosine with both Palmitoyl-GHK and GEKG
at a weight ratio of 4 parts Octanoyl Carnosine, 1 part
Palmitoyl-GHK, and 5 parts GEKG stimulates synthesis of hyaluronic
acid in a synergistic manner. Specifically, a combination of 100
ppm (0.01%) Octanoyl Carnosine, 25 ppm (0.0025%) Palmitoyl-GHK and
125 ppm (0.0125%) GEKG significantly stimulates the synthesis of
hyaluronic acid by 64%. However, Octanoyl Carnosine alone at same
concentration (100 pm) did not stimulate the synthesis of
hyaluronic acid (the results show a -5% increase, which, in fact,
means a non-significant inhibition of hyaluronic acid synthesis);
Palmitoyl-GHK alone (at 10 or 30 ppm) non-significantly stimulated
hyaluronic acid synthesis by only up to 11%; and GEKG alone (at 100
or 300 ppm) significantly stimulated hyaluronic acid synthesis but
only by up to 25%.
[0415] Consequently, if the effect of the combination of these
three peptides (or any derivatives thereof) was additive, one would
only expect only about a 30-40% increase in hyaluronic acid
synthesis (corresponding approximately to the sum of -5% plus 11%
plus 25%). However, unexpectedly, the combination of Octanoyl
Carnosine, Palmitoyl-GHK and GEKG at the weight ratio of 4:1:5
resulted in a stimulation hyaluronic acid formation by 64%, which
demonstrates that a combination of Octanoyl Carnosine,
Palmitoyl-GHK and GEKG at the weight ratio of 4:1:5 provides
synergistic effects (i.e., total effect of the combination of the
three peptides is greater than the sum of the effects of the three
peptides alone) in the stimulation of hyaluronic acid
formation.
[0416] Therefore, compositions containing Octanoyl Carnosine
combined with Palmitoyl-GHK and GEKG (or one or more derivatives
thereof) at the weight ratio of 4:1:5 according to the present
invention have therefore great potential in the field of cosmetics,
dermatology, wound healing, and any other area in the need of
treatments for conditions, disorders and diseases where hyaluronic
acid is altered. In addition, such compositions can be used for
maintaining healthy skin, skin rejuvenation, scarless wound
healing, restoration of damaged skin and mucosa, as well as for the
treatment of atrophy of any human tissue including but not limited
to vulvovaginal atrophy.
Example 6
Results of Clinical Tests on Examples According to the
Invention
[0417] The following clinical tests with compositions according to
the present invention illustrate a series of examples of the
different uses of the compositions and demonstrate the suitability
of the compositions for topical application in the field of
cosmetics, dermatology, wound healing, and any other areas in need
of treatments for conditions, disorders and diseases where
extracellular matrix components (including, but not limited to,
collagen III) are altered; and in particular for maintaining
healthy skin, skin rejuvenation, scarless wound healing,
restoration of damaged skin and mucosa, as well as for the
treatment of atrophy of any human tissue including but not limited
to vulvovaginal atrophy.
Clinical Test 1: Anti-Aging Study
[0418] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
(Composition D2; prepared as described in Example 4) was studied in
humans for maintaining healthy skin and skin rejuvenation. The
human subjects applied the cream twice daily to their face over a
period of six months. They were asked to apply the cream as they
would normally use a cosmetic facial product (e.g., facial
moisturizer). Evaluations for safety and effectiveness were
performed before the treatment with the cream started (baseline),
and after 1, 2, 3 and 6 months, respectively. The evaluations
included the assessment of the subject's face by trained
investigators for fine lines and coarse wrinkles in the pen-orbital
area, fine lines and coarse wrinkles in the pen-oral area, tactile
roughness, mottled hyperpigmentation, redness and telangiectasia,
skin pores and pore size, skin tone, and for global skin damage
using visual severity scores. The evaluations for effectiveness
further included clinical photography of the face (under
standardized conditions including position of the subject and
lighting), quantification of the skin roughness and wrinkle depths
of the peri-orbital skin area using the PRIMOS-3D System (GF
Messtechnik GmbH, Germany; utilizing standardized measuring and
lighting conditions in the `Overlay` mode), as well as the
measurement of skin elasticity of the upper cheek area with a
Cutometer.RTM. MPA580 device (Courage & Khazaka, Koln, Germany;
probe aperture 2 mm) after the subjects have been in a seated
position in the environmentally-controlled room for at least 30
minutes. The assessment of erythema (redness), edema (swelling),
dryness, and peeling (desquamation) was performed to determine the
safety of the cream. For this purpose, the investigator used the
following 5-point scale: 0=none, 1=mild, 2=moderate, 3=marked,
4=severe. Furthermore, adverse events were recorded during the
entire study duration. In a subset of subjects, a 3 mm punch biopsy
was taken from the pre-auricular skin area under local anesthesia
(i.e., up to 1 cc lidocaine with epinephrine given intradermally)
performed by the study doctor before treatment and at the end of
the six month treatment period. The sequential biopsies were nearly
adjacent to each other. The biopsy specimens were fixed in
formaldehyde, embedded in standard embedding medium, and then
evaluated for changes in epidermal and dermal structure (and for
changes of the extracellular matrix in particular) by histology. In
addition, the subjects reported about diverse attributes of the
cream (benefits, improvements, tolerability, cosmetic feel,
satisfaction, etc.) using questionnaires.
[0419] The following results were obtained in a study which
included females of skin types I to III with at least moderate
signs of facial wrinkles. Peri-orbital wrinkles improved (by at
least 1 unit) in 35% of the subjects after 1 month (from 2.9.+-.0.5
before treatment to 2.6.+-.0.6; mean.+-.SD, n=20 subjects), 71%
after 3 months (2.1.+-.0.6; n=17), and 88% after 6 months
(1.8.+-.0.4; n=17). Peri-oral wrinkles improved (by at least 1
unit) in 30% of the subjects after 1 month (from 2.7.+-.0.6 before
treatment to 2.4.+-.0.6; mean.+-.SD, n=20), 47% after 3 months
(2.2.+-.0.5; n=17), and 71% after 6 months (1.7.+-.0.5; n=17). The
product was well tolerated.
[0420] This study clearly demonstrated that an oil-in-water cream
containing 0.01% of the combination of Octanoyl Carnosine,
Palmitoyl-GHK and GEKG at a weight ratio 4:1:5 rejuvenates aged
skin. The composition was further well tolerated in all subjects
and did not cause any adverse events.
Clinical Test 2: Skin Restoration Studies after Cosmetic and
Dermatological Procedures
[0421] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
(Composition E; prepared as described in Example 4) was studied in
humans for enhancing the restoration of skin after cosmetic and
dermatological procedures such as chemical skin peels, skin
abrasion treatments, skin laser treatments, skin light treatments,
skin radiofrequency treatments, skin ultrasound, cold or hot
treatments of skin, non-surgical face lifts; or combinations
thereof. The subjects were asked to apply the cream once to
six-times daily to the affected skin area until at least the skin
has healed. This study involving different clinical case trials
demonstrated that an oil-in-water cream containing 0.01% of the
combination of Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a
weight ratio 4:1:5 enhances the restoration process of skin (e.g.,
decrease of time to heal, reduction of severity of side effects due
to procedure, reduction of skin inflammation, decrease of time for
restoring normal skin barrier, decrease of time for
re-epithelialization, increase of hydration, decrease of time to
appear normal again, help to improve appearance, help to reduce
signs of skin aging, contribution to increase collagen III
formation, does not result in scar formation, etc.) after cosmetic
and dermatological procedures. The composition was further well
tolerated.
[0422] A serum containing 0.5% of Octanoyl Carnosine (Composition
A3; prepared as described in Example 4) was studied in humans for
enhancing the restoration of skin after cosmetic and dermatological
procedures. The subjects were asked to apply the serum once to
six-times daily to the affected skin area until at least the skin
has healed. This study involving different clinical case trials
demonstrated that a serum containing 0.5% of Octanoyl Carnosine
enhances the restoration process of skin after cosmetic and
dermatological procedures. The composition was further well
tolerated.
Clinical Test 3: Wound Healing Studies
[0423] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
(Composition D2; prepared as described in Example 4) was studied in
humans for wound healing of surgical wounds, accidental wounds,
skin ulcers, and skin burns. The subjects were asked to apply the
cream once to six-times daily to the wounded area until at least
the wound has healed. This study involving different clinical case
trials demonstrated that an oil-in-water cream containing 0.01% of
the combination of Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a
weight ratio 4:1:5 accelerates wound closure and helps skin heal
without scar formation. The composition was further well
tolerated.
[0424] A serum containing 0.5% of Octanoyl Carnosine (Composition
A3; prepared as described in Example 4) was studied in humans for
wound healing of surgical wounds, accidental wounds, skin ulcers,
and skin burns. The subjects were asked to apply the serum once to
six-times daily to the wounded area until at least the wound has
healed. This study involving different clinical case trials
demonstrated that a serum containing 0.5% of Octanoyl Carnosine
accelerates wound closure and helps skin heal without scar
formation. The composition was further well tolerated.
Clinical Test 4: Skin Atrophy Studies
[0425] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
(Composition E; prepared as described in Example 4) was studied in
humans with skin atrophy due to long-term topical corticosteroids
use, or respectively, after menopause in women. The subjects were
asked to apply the cream once to six-times daily for at least four
weeks to the affected skin area. This study involving different
clinical case trials demonstrated that an oil-in-water cream
containing 0.01% of the combination of Octanoyl Carnosine,
Palmitoyl-GHK and GEKG at a weight ratio 4:1:5 reduce the severity
of skin atrophy and helps restore a normal skin barrier. The
composition was further well tolerated.
[0426] A serum containing 0.5% of Octanoyl Carnosine (Composition
A3; prepared as described in Example 4) was studied in humans with
skin atrophy due to long-term topical corticosteroids use, or
respectively, after menopause in women. The subjects were asked to
apply the serum once to six-times daily for at least four weeks to
the affected skin area. This study involving different clinical
case trials demonstrated that a serum containing 0.5% of Octanoyl
Carnosine reduces the severity of skin atrophy and helps restore a
normal skin barrier. The composition was further well
tolerated.
Clinical Test 5: Vulvovaginal Atrophy Studies
[0427] An oil-in-water cream containing 0.01% of the combination of
Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a weight ratio 4:1:5
(Composition K; prepared as described in Example 4) was studied in
women with vulvovaginal atrophy. In those women, vulvovaginal
atrophy was due to menopause (either naturally occurring, or
surgically induced), use of oral contraceptives, when undergoing
breast cancer treatments (e.g., including taking aromatase
inhibitors), or due to other causes altering their human sex
hormone levels. The women were asked to apply the cream once to
four-times daily for at least four weeks to their vulva by using
their index finger. This study involving different clinical case
trials demonstrated that an oil-in-water cream containing 0.01% of
the combination of Octanoyl Carnosine, Palmitoyl-GHK and GEKG at a
weight ratio 4:1:5 reduces the severity of the symptoms associated
with vulvovaginal atrophy (e.g., dryness of vulva, feel of rawness,
discomfort, pain with sex, itch, etc.) as well as reduces the
severity of vulvovaginal atrophy; in particular for the vulva.
Clinical Test 6: Studies in Other Conditions, Disorders and
Diseases where Extracellular Matrix Components are Altered
[0428] Compositions according to the present invention were also
studied for the treatment of other conditions, disorders and
diseases where extracellular matrix components are altered
including atopic dermatitis, eczema, scars and keloids, atrophie
blanche, vulvar lichen sclerosus, epidermolysis bullosa,
Ehlers-Danlos syndromes, and the Marfan syndrome. The subjects were
asked to apply the compositions between once to six-times daily for
at least four weeks to the affected skin area. This study involving
different clinical case trials demonstrated that the compositions
help reduce the symptoms and the severity of those conditions,
disorders and diseases. The composition was generally well
tolerated.
EQUIVALENTS
[0429] The details of one or more embodiments of the invention are
set forth in the accompanying description above. Although any
methods and materials similar or equivalent to those described
herein can be used in the practice or testing of the present
invention, the preferred methods and materials are now described.
Other features, objects, and advantages of the invention will be
apparent from the description and from the claims. In the
specification and the appended claims, the singular forms include
plural referents unless the context clearly dictates otherwise.
Unless defined otherwise, all technical and scientific terms used
herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. All
patents and publications cited in this specification are
incorporated by reference.
[0430] The foregoing description has been presented only for the
purposes of illustration and is not intended to limit the invention
to the precise form disclosed, but by the claims appended hereto.
Sequence CWU 1
1
1314PRTArtificial SequenceSynthesized peptide 1Gly Glu Lys Gly 1
24PRTArtificial SequenceSynthesized peptide 2Gly Xaa Xaa Gly 1
34PRTArtificial SequenceSynthesized peptide 3Pro Xaa Xaa Pro 1
44PRTArtificial SequenceSynthesized peptide 4Pro Xaa Xaa Lys 1
54PRTArtificial SequenceSynthesized peptide 5Gly Glu Pro Gly 1
64PRTArtificial SequenceSynthesized peptide 6Gly Pro Pro Gly 1
74PRTArtificial SequenceSynthesized peptide 7Pro Gly Pro Pro 1
84PRTArtificial SequenceSynthesized peptide 8Pro Lys Glu Lys 1
94PRTArtificial SequenceSynthesized peptide 9Gly Gln Pro Arg 1
104PRTArtificial SequenceSynthesized peptide 10Arg Ser Arg Lys 1
115PRTArtificial SequenceSynthesized peptide 11Lys Thr Thr Lys Ser
1 5 126PRTArtificial SequenceSynthesized peptide 12Val Gly Val Ala
Pro Gly 1 5 135PRTArtificial SequenceSynthesized peptide 13Tyr Gly
Gly Phe Xaa 1 5
* * * * *